biofiles 3

Chondroitin sulfate proteoglycans modulate secreted chemotactic proteins that regulate axon guidance in neural cell development. These guidance proteins have distinct binding specificities for sulfate groups (sulfur shown in red) within these glycosaminoglycan chains.

Volume 3, Number 10

BioFilesLife Science

GlycobiologyGlycosaminoglycans and Polysaccharides

Glycosaminoglycan Sulfation and Signaling

Dextrans and Related Polysaccharides

GlycoProfile™ b-Elimination Kit

2

Life Science

BioFilesVolume 3, Number 10

Table of Contents

Introduction .........................................................3

Glycosaminoglycan Sulfation and Signaling ......................................................4 Biosynthesis of Heparan/Heparin and Chondroitin/Dermatan .............................. 4 Heparan Sulfate Signaling .................................. 6 Chondroitin Sulfate/Dermatan Sulfate Signaling .............................................. 7 Research Techniques .......................................... 8 Glycosaminoglycans and Proteoglycans ........... 10 Glycosaminoglycan Degrading Enzymes .......... 13 Antibodies ........................................................ 14 Prestige Antibodies™ .................................. 14

Antibodies to PG Core Proteins ................... 15

Antibodies to GAG Synthesis/Modification Enzymes .................. 15

Additional Antibodies ................................. 15

Glycosaminoglycan Disaccharides .................... 16 Monosaccharide Sulfates ................................. 16

Dextran and Related Polysaccharides ...............17 Dextrans ........................................................... 19 Dextran ...................................................... 19

Dextran Sulfate ........................................... 22

Immobilized Dextran ................................... 22

Dextrins and Pullulans ..................................... 23 Additional Gel Permeation Chromatography (GPC) Standards and Sets ........................................ 23

GlycoProfile™ β-Elimination Kit ........................24 Related Kits for Deglycosylation ...................... 26

Technical Content: Vicki Caligur, B.Sc., Roland Wohlgemuth, Ph.D.

Your gateway to Biochemicals and Reagents for Life Science Research from Sigma®

BioFiles Online allows you to:

• Access any issue of BioFiles

• Subscribe for:

✓ future issues

✓ eBioFiles email notifications

• Stay up-to-date on the latest BioNews from Sigma

Register today for upcoming issues and eBioFiles announcements and receive a free IUBMB-Sigma-Nicholson Metabolic Pathway Chart: sigma.com/biofiles

Enzymes for Carbohydrate Analysis

The carbohydrate analysis resource from the Enzyme Explorer allows you to select from either the enzyme or carbohydrate to find detailed enzyme specificity diagrams and information for complex carbohydrates.

• Information on 25 enzymes and over 20 complex carbohydrates

• Quick links to products and references

• Complete list of enzymes for PTM analysis and carbohydrate analysis

Start using the carbohydrate analysis resource at sigma-aldrich.com/carboanalysis

Glycobiology Webinar

The GlycoProfile™ Metabolic Labeling Reagent Technology Webinar explains the GlycoProfile kits and reagents abilities to incorporate, express and detect unnatural glycoproteins within cells.

Applications covered in the webinar:

• Selectively targeting labeled sugars

• Monitoring differential presentation of important glycans

• Visualizing internally and externally labeled glycoproteins

• Purifying glycoproteins

View the webinar at sigma-aldrich.com/glycoprofile

http://www.sigma.com/biofiles

http://www.sigma-aldrich.com/carboanalysis

http://www.sigma-aldrich.com/glycoprofile

Our Innovation, Your Research — Shaping the Future of Life Science 3

Intro

du

ction

IntroductionVicki CaligurProduct Manager, Specialty [email protected]

Polysaccharides are structural components of plant, animal, and bacterial cells. As a group, polysaccharides, such as starch, glycogen, and cellulose, are ubiquitous and common. However, when specific classes of polysaccharides are examined, it is clear

they have great variation based on origin, structure, and function. In this Glycobiology issue of BioFiles, we have focused on two specific types of polysaccharides: glycosaminoglycans and dextran.

Even within the specific class of glycosaminoglycans, the diversity of function is so extensive that a comprehensive review is not possible. Glycosaminoglycans are primarily found in animal tissue, and their function was initially thought to be solely that of space-occupying components of the extracellular matrix. We now know that glycosaminoglycans contribute to the functionality of proteoglycans and participate in cellular signaling, neuron development, lipoprotein metabolism, inflammation, and bacterial infection.

The structural complexity of glycosaminoglycans is necessary for functionality but also makes structure confirmation extremely difficult, as demonstrated by the heparin contamination crisis in early 2008.

In the early 20th century, heparin was isolated and found to act as an anticoagulant, and it has been used medically for over 70 years. Low molecular weight heparin modified by various techniques has been developed for use as an anticoagulant to reduce heparin-induced thrombocytopenia. In 1983, the pentasaccharide sequence within heparin that possesses the ability to block factor Xa and antithrombin activity was identified.1 The pentasaccharide fondaparinux is now commercially synthesized and has been approved for medical use in the United States and Europe as an anticoagulant under the trade name ARIXTRA®. Although these alternatives are available, heparin from natural sources continues to be widely used clinically.

In the spring of 2008, at least 20 patient deaths were associated with contaminated heparin that was not detected by routine analysis. An international team of researchers identified the impurity as a variant of chondroitin sulfate, a different glycosaminoglycan found in cartilage, which had been modified with sulfate groups in an unnatural pattern. The conclusive identification required multiple techniques, including multidimensional NMR spectroscopy supported by enzymatic digestion and HPLC analysis and was validated by chemical synthesis of an oversulfated chondroitin that matched the unknown contaminant.2 As a result of this exhaustive analysis,

capillary electrophoresis and NMR spectroscopy are now required by the United States FDA to screen heparin for similar contamination.

Dextran, a glucose polymer isolated from the bacteria Leuconostoc mesenteroides was first applied to medical use as a plasma volume expander and an antithrombotic during World War II. The structural features and applications of dextran, dextran sulfate, and pullulan are reviewed in this issue.

Finally, the GlycoProfile™ β-Elimination Kit is Sigma®’s newest product for removing glycans from glycoproteins. Unlike other chemical deglycosylation techniques, β-elimination allows researchers to recover both the glycan pool and the protein core for downstream analysis and application. The GlycoProfile β-Elimination Kit allows for complete glycoproteomic analysis of O-linked glycoproteins as never before possible.

References: 1. Petitou, M., Casu, B., and Lindahl, U., 1976-1983, a critical period in the history of

heparin: the discovery of the antithrombin binding site. Biochimie, 85, 83-89 (2003).2. Guerrini, M., Beccati, D., Shriver, Z., Naggi, A., Viswanathan, K., Bisio, A., Capila, I.,

Lansing, J.C., Guglieri, S., Fraser, B., Al-Hakim, A., Gunay, N.S., Zhang, Z., Robinson, L., Buhse, L., Nasr, M., Woodcock, J., Langer, R., Venkataraman, G., Linhardt, R.J., Casu, B., Torri, G., and Sasisekharan, R. Oversulfated chondroitin sulfate is a contaminant in heparin associated with adverse clinical events. Nat. Biotechnol., 26, 669-75 (2008).

Please visit the BioBlog to

read about and comment on

discoveries and trends reported

at the 2008 Annual Conference

of the Society for Glycobiology.

http://www.sigmabioblogs.com

4 Order: sigma.com/order Technical service: sigma.com/techinfosigma.com/lifescience

Gly

cosa

min

og

lyca

n S

ulf

atio

n a

nd

Sig

nal

ing

Glycosaminoglycans are large linear polysaccharides constructed of repeating disaccharide units. The disaccharide units that are the basis of all glycosaminoglycans (except keratan) are a combination of an uronic acid (either glucuronic acid or iduronic acid) and an amino sugar (either N-acetyl-D-glucosamine or N-acetyl-D-galactosamine). There are four classifications of glycosaminoglycans:

� Hyaluronan

� Chondroitin sulfate/dermatan sulfate

� Heparan sulfate/heparin, and

� Keratan sulfate.

A given glycosaminoglycan will contain only one type of amino sugar, but may contain both uronic acid forms, as iduronic acid is produced by enzyme-induced epimerization of glucuronic acid. The core disaccharide unit of keratan is N-acetyl-D-lactosamine ([1→3]β-D-galactose-β[1→4]-N-acetyl-D-glucosamine; β-D-Gal-[1→4]GlclNAc-[1→3]) and the galactose residue is not available for epimerization.

Glycosaminoglycan structures have a high degree of heterogeneity (termed “fine structure”) with respect to molecular weight, disaccharide construction, and sulfation. This variability can be attributed to the fact that glycosaminoglycan synthesis is dynamic. While protein and nucleic acid syntheses are template-based processes, glycosaminoglycan synthesis has no template but is modulated by the processing enzymes present. In addition, glycosaminoglycans, with the exception of hyaluronan, may have sulfate groups added to specific locations of the disaccharide subunit. However, enzymatic sulfation and epimerization reactions do not go to completion, resulting in structural heterogeneity within the polysaccharide molecule. This heterogeneity and dynamic processing make investigation of the structure/function relationships of glycosaminoglycans challenging.

Proteoglycans are proteins that have one or more glycosaminoglycan polymers attached to the protein core via an O-linkage through a serine residue. Glycosaminoglycans, whether separately or as a component of a proteoglycan, can bind to and interact with proteins. These glycosaminoglycan-protein interactions have essential roles in cellular processes including proliferation, cellular differentiation and development, and there have been efforts to relate specific sulfation patterns with resultant processes.

Hyaluronan is the only glycosaminoglycan that is not sulfated and does not attach to proteins to form proteoglycans. It has been associated with CD44 binding and signaling. The key function of keratan sulfate is as a structural component of cornea proteoglycans and not signaling effects. The glycosaminoglycans primarily involved in cellular signaling are heparan sulfate (HS)/heparin and chondroitin sulfate (CS)/dermatan sulfate (DS). This discussion will focus on heparan/heparin and chondroitin/dermatan and how sulfation impacts protein binding interactions and signaling effects by those glycosaminoglycans.

Biosynthesis of Heparan/Heparin and Chondroitin/DermatanThe synthesis of heparan and chondroitin chains on a proteoglycan begins in the same way via an O-linked tetrasaccharide. The GAG chain synthesis is initiated by the attachment of xylose (Xyl) by xylosyltransferase (XylT) to a serine residue of the protein core. Two galactose (Gal) sugars are attached to the xylose sequentially by the enzymes galactosyltransferase I (GalT-I) and galactosyltransferase II (GalT-II). Glucuronyltransferase I (GlcAT-I) attaches the fourth sugar, glucuronic acid (GlcA) to the initial chain. At this point, the synthesis diverges (see Figure 1). A heparan polysaccharide continues with the attachment of an N-acetyl-D-glucosamine (GlcNAc) residue by N-acetylglucosaminyltransferase I. Alternatively, a chondroitin polysaccharide continues with the attachment of an N-acetyl-D-galactosamine (GalNAc) residue by N-acetyl galactos-aminyl transferase I.1,2

Ser

Ser

Ser

Ser Ser

Ser

Figure 1. Biosynthesis of heparan and chondroitin chains. After the core tetrasaccharide (Xyl-Gal-Gal-GlcA-) is synthesized, the polymer can become a heparan chain by the addition of GlcNAc (shown left) or a chondroitin chain by the addition of GalNAc (shown right).

For heparan, the glycosaminoglycan is polymerized by the alternating addition of glucuronic acid and N-acetyl-D-glucosamine residues, to a final length of up to 200 disaccharides. The critical glycosyltransferases for heparan polymerization are exostosin 1 (EXT1) and exostosin 2 (EXT2). Modification of the polysaccharide is necessary for development of the heterogeneous fine structure. Heparan is modified by the N-deacetylase/N-sulfotransferase enzymes NDST1 and NDST2, which replace the N-acetyl groups (GlcNAc) with N-sulfate groups (GlcNS) on a glucosamine residue. The glucuronic acid units of the heparan chain are also susceptible to modification. Glucuronic acid (GlcA) may be converted to its epimer iduronic acid by the enzyme glucuronyl C5-epimerase. The iduronic acid moiety (IdoA) may be further modified by sulfation at the 2-O-position (IdoA-2S) through the activity of the enzyme heparan sulfate 2-O-sulfotransferase (HS2ST).2,3

Glycosaminoglycan Sulfation and Signaling

http://www.sigma.com/order

http://www.sigma.com/techinfo


Glyco

samin

og

lycan Su

lfation

and

Sign

aling

These disaccharides may also be sulfated at the C-6 position of glucosamine (see Figure 2). Three variants of the heparan sulfate 6-O-sulfotransferase gene (HS6ST) have been identified.4 The uncommon form of sulfation is at the 3-O-position of the glucosamine ring and is mediated by the enzyme heparan sulfate 3-O-sulfotransferase (HS3ST). There are six isoforms of the HS3ST enzyme (1, 2, 3A, 3B, 4, and 5). Both the HS6ST and HS3ST enzymes have specific substrate preferences.

6S 6S

6S 6S6S

Ser

Ser

Ser

Ser

6S Ser

Ser

NDST1, NDST2

HS2ST

HS3ST (1,2,3A,3B,4,5)

HS6ST (1,2,3)

Heparin Antithrombin III Pentasaccharide

2S

2S

2S

6S

3SNS

NSNS NS NS

NSNS NS NS

NSNS NS NS

NSNS NS NS

NS NS NS

Glucuronyl C5-epimerase

Figure 2. The modification of the heparan core is caused by a cascade of enzymes. The resultant heparan sulfate contains IdoA residues and domains with a high degree of sulfation. The pentasaccharide antithrombin II sequence associated with heparin is shown in the final image. See text for further explanation.

Sulfation is concentrated in specific regions of the heparan polysaccharide. As a result, heparan sulfate (HS) has regions of low sulfation interspersed with regions of high sulfation, with transitional regions separating the two. Highly sulfated domains, termed “composite sulfated regions” or S-domains, contain between 2 and 7 or 8 sequential GlcNS-IdoA-2S disaccharides. Other regions of the heparan polymer are largely unmodified and remain acetylated, constructed of GlcNAc-GlcA disaccharide units. Transition zones connect the two domains (see Figure 3). The fine structure complexity of HS is caused by both the variation in the disaccharide units and the overall variability created by the domain structure.5,6

S-Domain S-Domain S-Domain

N-Acetylated Domains

Figure 3. Heparan sulfate has regions of high sulfation (S-domains) interspersed with less modified regions (N-acetylated domains). A transition zone connects the two regions.

Heparin is a specific member of the heparan sulfate family of glycosaminoglycans. Heparin is expressed exclusively by mast cells, including mucosa mast cells, and heparin isolated from porcine intestinal cells has been approved by medical regulatory agencies for use as an anticoagulant. This anticoagulant activity of heparin has been shown to be due to the specific pentasaccharide (five-sugar) structure (see Figure 4).

6S 6S

2S3SNS

6S

NS

Figure 4. The pentasaccharide antithrombin binding site of heparin. Sites of sulfation are indicated.

This sequence contains a rare 3-O-sulfated glucosamine and is a specific ligand for the protease inhibitor antithrombin. Heparin binding of antithrombin blocks the activation of factors of the coagulation cascade mechanism. While heparin is frequently referred to in conjunction with heparan sulfate, it should be considered as a unique member of the heparan sulfate family and not interchangeable with heparan sulfate.2,7

For chondroitin synthesis, the glycosaminoglycan is polymerized by the alternating addition of glucuronic acid and N-acetyl-D-galactosamine residues. The completed polymer typically has 40 to over 100 disaccharide units. Modification of the chondroitin structure is also dynamic, generating fine structures that have type classifications that have not been applied to heparan. Chondroitins have been classified into 4 major types, based on location of sulfation on the disaccharide:

� Chondroitin sulfate A (CS-A) GlcA-GalNAc-4-SO4 (ΔDi-4S; A)

� Chondroitin sulfate C (CS-C) GlcA-GalNAc-6-SO4 (ΔDi-6S; C)

� Chondroitin sulfate D (CS-D) GlcA-2-SO4-GalNAc-6-SO4 (ΔDi-diSD; D)

� Chondroitin sulfate E (CS-E) GlcA-GalNAc-4,6-diSO4 (ΔDi-diSE; E)

Less common chondroitin disaccharides B, H, and K have also been identified.

Key to Monosaccharide Symbols

β-D-Glucose (Glc) β-D-Xylose (Xyl)

β-D-Mannose (Man)α-N-Acetylneuraminic acid;

Sialic acid (NeuNAc)

β-D-Galactose (Gal) β-D-Glucuronic acid (GlcA)

β-D-N-Acetyl glucosamine (GlcNAc) α-L-Iduronic acid (IdoA)

β-D-N-Acetyl galactosamine (GalNAc) α-L-Fucose (Fuc)


Gly

cosa

min

og

lyca

n S

ulf

atio

n a

nd

Sig

nal

ing

Dermatan, also known as chondroitin sulfate B, undergoes significant epimerization of glucuronic acid (GlcA) to iduronic acid (IdoA). The enzyme 2-sulfotransferase preferentially sulfates IdoA adjacent to GalNAc-4-SO4. As a result of this enzymatic specificity, the primary disaccharide unit of dermatan sulfate is IdoA-2-SO4-GalNAc-4-SO4 (iB). Other rare dermatan sulfates have been recognized, and are named similarly to their chondroitin sulfate counterparts (iA, iC, iD, and iE), using the letter “i” to indicate the iduronic acid residue.

As the sulfation and epimerization reactions occur to varying degrees, there is no absolute structure of any chondroitin sulfate/dermatan sulfate, and chondroitin/dermatan contains many fine structures much like heparan sulfate.1

Heparan Sulfate SignalingHeparan sulfate proteoglycans (HSPGs) have been extensively studied for their role in extracellular signaling. HSPGs interact with and regulate growth factors, chemokines, and cytokines. Signaling pathways for apoptosis, cellular development, and adhesion are regulated by proteoglycans. Genetic defects in heparan synthesis and modifying enzymes may affect growth factor signaling and produce morphological defects.8 Drosophila mutants and gene knockout mouse models have been used to eliminate critical heparan synthesis and modification enzymes, and subsequently identify the developmental processes disrupted. Experiments that use Drosophila containing mutations of the protein core of proteoglycans have been performed and have shown that the protein core also affects development. It has been theorized that both the HS and the protein core have roles in the mediation of growth factor binding and cell signaling.6

HS and HSPGs participate in fibroblast growth factor-2 (FGF-2) activation in response to injury and inflammation. After injury, HSs break free from the cell membrane or their proteoglycan serine linkage and become soluble. Heparanase that is released by cells involved in inflammation cleaves the released heparan sulfate into smaller heparin-like oligosaccharides. While intact HS does not activate FGF-2, these smaller oligosaccharides are able to activate FGF-2. This processing of free HS by heparanase in response to inflammation has been theorized to be a necessary step to wound healing.2,9

The binding of HSPGs to FGF-2 has been studied as well for a possible association with malignant transformation of breast carcinoma cells by FGF-2 in vitro. Fibroblast growth factors require the presence of heparan sulfate or heparin to stabilize binding with tyrosine kinase signaling receptors (FGFR-1 through FGFR-4). Mundhenke found that HSPGs isolated from the MCF-7 breast-carcinoma cell line had an elevated ability to promote formation of the FGF-2/HSPG/FGFR-1 complex as compared to normal epithelial cells. HSPGs have also been shown to affect signaling by attachment to other growth factors including FGF-7.10

Using glycosaminoglycan microarrays, Shipp and Hsieh-Wilson showed that individual members of the FGF family require specific sulfation patterns for HS binding. In addition to confirming sulfation specificities previously reported for FGF-1, FGF-2, FGF-4, they were able to characterize the heparan sulfation requirements

for FGF-16 and FGF-17. Using the microarrays, chemotactic proteins, including slit2, that regulate axon extension were found to have preferential binding to specific sulfation patterns. This finding was validated by co-culturing embryonic olfactory bulb cells with HEK293 cells expressing slit2. The cultures were supplemented with heparan sulfate variants containing different sulfation patterns. After staining with anti-τ antibody for axon visualization, the cultures showed that highly sulfated heparan sulfate reduced slit2-mediated axon repulsion, while desulfated heparin or N-acetylated heparin had no effect on slit2 repulsion.11

The genes for the heparan polymerases exostosin 1 (EXT1) and exostosin 2 (EXT2) were initially thought to be tumor suppressor genes. Mutations of EXT-type genes in Drosophila reduced the level of heparan synthesis and inhibit hedgehog (Hh), Wingless (Wg, which correlates to Wnt), and Dpp signaling pathways, indicating that HS participates in the signaling function of these pathways.12,13 The protein core of the proteoglycan has been shown to contribute to HSPG signaling effects as well. Wnt signaling was modulated by the processed protein core of the HSPG glypican 3, with the implication that proper signaling requires contributions by both the HS and the core protein components.14

A similar protein-glycosaminoglycan synergy toward binding affinity was identified for the association of interleukin-8 (IL-8) with the HSPG syndecan 2 on human umbilical vein endothelial cells. IL-8 is a chemokine that is secreted at sites of inflammation by cytokine activated endothelial cells and which is retained on the cell surface by interactions with HSPGs. While glycosylated syndecan 2 showed strong affinity for IL-8, the nonglycosylated protein core also demonstrated weak IL-8 binding.15

CD44 (Pgp-1, ECM-III, H-CAM) functions as a homing receptor and is most commonly the cellular receptor for hyaluronic acid. Jones, et al., found that some CD44 isoforms exist as heparan sulfate proteoglycans, and that CD44/HSPG was able to bind FGF-2, vascular endothelial growth factor (VEGF), and heparin-binding epidermal growth factor. CD44/HSPG isoforms were overexpressed by macrophages within inflamed rheumatoid arthritis tissue but minimally expressed in noninflamed osteoarthritis tissue. Based on the experimental data, it was theorized that CD44/HSPG isoforms are involved in the regulation of growth factor activity in inflammatory arthritis.16

You, et al., reported that the tumor necrosis factor receptor family member Decoy receptor 3 (DcR3) could induce apoptosis in dendritic cells (DC) through binding between DcR3 and DC surface heparan sulfate proteoglycans This was based on the observation that HBD.Fc, the recombinant protein containing both the heparan sulfate-binding domain (HBD) of DcR3 and the Fc portion of human IgG1, functions in the same manner as DcR3.Fc to induce apoptosis in dendritic cells.17

In addition to sulfate modifications present on the heparan chain, cell surface sulfatases are critical to growth factor signaling. The human sulfatases Sulf1 and Sulf2 are 6-O-endosulfatases and are able to remove 6-O-sulfate from the highly sulfated regions of the polysaccharide.6 The association between 6-O-sulfation of HS and FGF signaling has been studied from many aspects. In a review by Nakato and Kimata, the authors discussed how the fine structures




Glyco

samin

og

lycan Su

lfation

and

Sign

aling

of HS, especially those containing 6-O-sulfation, modulate the growth factor signaling functions of HSPGs.8 Increased Sulf-1 expression by the breast cancer cell line MDA-MB-468 inhibited angiogenesis and cancer cell proliferation in vivo. The inhibition was attributed in part to the reduced ability of the vascular HS to form a stable FGF-2/HSPG/FGFR-1 complex. The inference is that Sulf-1 overexpression reduces the degree of 6-O-sulfation in HS fragments, and that 6-O-sulfation contributes to the stabilization of the FGF-2/HSPG/FGFR-1 complex.18 More recently, experiments with murine embryonic fibroblasts (MEF) HS-6-O-transferase deficient mice (HS6ST-1-KO, HS6ST-2-KO, and double knock-out) showed that FGF-2 and FGF-4-dependent signaling in MEF was significantly reduced compared to that of wild-type fibroblasts.19

The vascular endothelial growth factor VEGF165 contains a heparin-binding domain that is only weakly activated by 2-O-desulfated heparin or 6-O-desulfated heparin.20 6-O-sulfation has also been shown to affect the signaling pathways of Wnt and bone morphogenic protein (BMP). It has been suggested that there is a dynamic interaction between the Sulf enzymes and HS biosynthetic enzymes, rather than independent processes.6

Chondroitin Sulfate/Dermatan Sulfate SignalingWhile HS has been of primary interest in glycosaminoglycan signaling effects, chondroitin sulfate (CS) and dermatan sulfate (DS) have also been recognized as affecting signaling pathways,21 and that these interactions are dependent on the polysaccharide fine structure. Both the glycosaminoglycan components and the proteoglycan have been investigated for contributions to biological function. Because of the fine structure and heterogenetity, both CS and DS units may exist as hybrids.22 Research has primarily focused on the contribution by the specific chondroitin suflate proteoglycans (CSPGs) and dermatan sulfate proteoglycans (DSPGs) aggrecan, decorin, biglycan, versican and the syndecans. Additional CSPGs/DSPGs of interest are neurocan, brevican, bamacan, betaglycan, serglycin, endocan, and appican.

In much the same manner as HS, CS/DS and CSPGs /DSPGs participate in fibroblast growth factor activation in response to injury and inflammation.2,22 After injury, the glycosaminoglycan chains are cleaved at their serine linkage, or the proteoglycans separate from the cell membrane and become soluble. Soluble dermatan sulfate is able to activate FGF-2 23 and FGF-7, initiating cellular proliferation via FGF-7 through the mitogen-activated protein-kinase (MAPK) pathway.24 The minimum dermatan sulfate structure required by FGF-2 is an octasaccharide and the requirement by FGF-7 is a decasaccharide. 4-O-Sulfation is also required for FGF-dependent cell proliferation, while additional 2-O-sulfation or 6-O-sulfation did not increase the disaccharide activity.25

The hepatic growth factor/scatter factor (HGF/SF) is known to participate in morphogenesis and cellular differentiation, organogenesis, and angiogenesis. Endocan, a DSPG that is regulated by proinflammatory cytokines, requires the presence of the glycosaminoglycan moiety in order to promote HGF/SF-induced proliferation of human embryonic kidney cells.26 HGF/SF binding demonstrated preferential binding by 2,6-O-disulfated DS and highly sulfated HS, but not CS, indicating a selective requirement for either iduronic acid residues or a high degree of sulfation, although highly sulfated polysaccharides were less bound than the iduronic acid-containing glycosaminoglycans.27

While the fine structure of CS/DS affects binding and signaling effects, it turns out that growth factors can modulate the expression and structure of the CS/DS glycosaminoglycan. Cultured human lung fibroblasts were treated with transforming growth factor-β1 (TGF-β1) alone, or in combination with epidermal growth factor and platelet-derived growth factor. Cells treated with TGF-β1 or the combination of growth factors increased their expression of CS/DS proteoglycans. In addition, the sulfation and epimerization profiles of the chondroitin/dermatan chains were significantly modified compared to untreated fibroblasts.28

Tailored for the life science researcher, BioFiles aligns our vast array of products within a relevant research topic.

Life Science Innovations piques your interest with examples

of new and emerging technologies put forth in a

fresh, unique way that applies to your area of study.

sigma-aldrich.com

Life Science Innovations and BioFiles offer collaboration and innovation from our scientists to you.

Visit Life Science Innovations Online at sigma.com/innovations

A Perfect Fit!

http://www.sigma-aldrich.com/innovations


Gly

cosa

min

og

lyca

n S

ulf

atio

n a

nd

Sig

nal

ing

The highly sulfated CS-E type has been shown to bind to heparin binding growth factors midkine (MK), pleiotropin (PTN), heparin-binding epidermal growth factor-like growth factor (HB-EGF), FGF-16, and FGF-18. As many of these growth factors are expressed in the mammalian brain, it was proposed that CS-E and CSPGs are critical to the development of the brain and central nervous system.29 Subsequent work using an antibody specific for both the CS-E disaccharide and the oversulfated DS-E disaccharide (IdoA-GalNAc-4,6-diSO4; iE) revealed the presence of these glycosaminoglycan sequences in the mouse developing brain. The strong expression of the gene for GalNAc4S-6ST was associated with brain development, and demonstrated that E/iE-containing CS/DS chains are critical in brain development with the implication that E/iE-containing CS/DS chains participate in neurogenesis, axon guidance, and/or neuronal survival.30

While the glycosaminoglycan sequence and sulfation pattern regulate signaling, the protein core of the glycosaminoglycan also contributes to the signaling ability. The ligand binding of the CS/DS proteoglycans decorin and biglycan to transforming growth factor-β (TGF-β), tumor necrosis factor-β (TNF-β), and Wnt-induced secreted protein-1 (WISP1) is mediated by both the DS component and the protein cores. A recent review by Seidler and Dreier analyzed both the galactosaminoglycan component and the protein core of decorin as critical to the ability of decorin to function as a ligand for other signaling molecules.31

The enzymes N-acetylgalactosamine-4-sulfatase (arylsulfatase B; ASB) and galactose-6-sulfatase (GALNS) hydrolyze sulfate groups of CS/DS. MCS-7 cells that overexpressed ASB or GALNS produced increased expression of CS/DS proteoglycans syndecan-1 and decorin. Alernatively, when siRNA was used to silence ASB or GALNS expression in MCS-7 cells, the level of CS content increased and syndecan-1 expression was inhibited. The results demonstrate a role for these sulfatases in CS development and the regulation of proteoglycan expression.32

Research TechniquesA variety of experimental techniques have been employed to identify the structure of the glycosaminoglycan chain, the degree and location of sulfation, and the relationship between polysaccharide component and protein core. Knock out mice that lack a designated heparan or chondroitin/dermatan synthesis enzyme have been used to help identify the pathways affected by the lack of glycosaminoglycan modification. Table 1 lists several of the enzymes involved in glycosaminoglycan synthesis and modification and their genes. Table 2 lists the genes for proteoglycan core protein expression. RNA interference has been applied in vitro as another means to eliminate synthesis and modification enzymes to study downstream cellular processes. ELISA analysis with proteoglycan core specific antibodies has been used to monitor protein expression, while quantitative RT-PCR has been applied to study gene regulation.

Genes of Heparan and Chondroitin Biosynthesis Enzymes

Gene Enzyme Name

ARSB Arylsulfatase B (N-Acetylgalactosamine-4-sulfatase )

C4ST1 (CHST11) Chondroitin 4-O-sulfotransferase 1



ChGn Chondroitin β1,4 N-acetylgalactosaminyltransferase

CHST1 Carbohydrate (chondroitin 6/keratan) sulfotransferase 1

CHST2 Carbohydrate (chondroitin 6/keratan) sulfotransferase 2

CHST3 Carbohydrate (chondroitin 6) sulfotransferase 3

CHST7 (C6ST-2) Chondroitin 6-sulfotransferase 2

CHSY1 Chondroitin synthase (Carbohydrate synthase 1)

D4ST1 (CHST14) Dermatan 4-sulfotransferase 1

DSE Dermatan sulfate epimerase

EXT1 Exostosin 1 (N-Acetylglucosaminyl-proteoglycan 4-β-glucuronosyltransferase)

EXT2 Exostosin 2(N-Acetylglucosaminyl-proteoglycan 4-β-glucuronosyltransferase)

GALNS Galactosamine (N-acetyl)-6-sulfate sulfatase (N-Acetylgalactosamine-6-sulfatase)

GLCE Glucuronic acid epimerase(Heparin/heparan sulfate glucuronic acid C5 epimerase)

HS2ST1 Heparan sulfate 2-O-sulfotransferase I



NDST1 N-Deacetylase/N-sulfotransferase 1



SULF1 Sulfatase 1

SULF2 Sulfatase 2

Table 1. Genes of heparan and chondroitin biosynthesis enzymes. MISSION® siRNA and/or shRNA are available for all genes listed.

Genes of Proteoglycan Core Proteins

Gene Protein Name

ACAN Aggrecan

AGRN Agrin

APP Amyloid β (A4) precursor protein (Appican)

BGN Biglycan

DCN Decorin

GPC1 Glypican 1

GPC2 Glypican 2

GPC3 Glypican 3

GPC4 Glypican 4

GPC5 Glypican 5

GPC6 Glypican 6

HSPG2 (PLC) Perlecan

SDC1 Syndecan 1

SDC2 Syndecan 2

SDC3 Syndecan 3

SDC4 Syndecan 4

SRGN Serglycin

Table 2. Genes of proteoglycan core proteins. MISSION® siRNA and/or shRNA are available for all genes listed.




Glyco

samin

og

lycan Su

lfation

and

Sign

aling

Enzymatic degradation is often used to cleave glycosaminoglycans into disaccharide units for subsequent analysis. Amide-hydrophilic interaction chromatography (HILIC) liquid chromatography with tandem mass spectrometry (LC/MS/MS) has been applied for comparative glycomics of chondroitin sulfate isoforms. As carbohydrates are inherently difficult to detect by HPLC, they are often modified by reductive amination to covalently attach a detection molecule, typically 2-aminobenzoic acid (2-AB), 2-anthranilic acid (2-AA) or 2-aminopyridine (2-AP). Stable isotopically labeled 2-anthranilic acid was used to comparatively profile CS disaccharides from different tissues.33

Poly-L-lysine coated slides were used to create microarrays of desulfated HS glycosaminoglycans. The microarrays were used to profile the binding of FGF-2 and other FGF factors against different sulfation patterns. The microarray technique provides a high throughput method to study protein-glycosaminoglycan binding.11

High-resolution NMR spectroscopy, X-ray crystallography, and molecular modeling are valuable methods applied to the investigation of glycosaminoglycan-protein structural interactions. Some of the resulting glycan-protein data has been recorded and is publicly available from the Centre de Recherches sur les Macromolécules Végétales (CER-MAV; http://www.cermav.cnrs.fr/glyco3d).34

A glycomics approach has been used to monitor the regulation of glycosaminoglycan biosynthesis during differentiation of mouse embryonic stem cells. Glycosaminoglycan content and composition changed during cellular transition from stem cells to embryoid bodies and extraembryonic endodermal cells, with 4 to 6-fold increases in CS/DS synthesis and 5 to 8-fold increases in heparan sulfate synthesis. mRNA transcription levels for glycosaminoglycan synthesis and modifying enzymes were unchanged and did not appear to correlate to the changes in glycosaminoglycan levels found during differentiation.35

SummaryMuch has been learned in the last few years about glycosaminoglycans. The synthesis mechanisms have been elucidated in great detail and their contribution to proteoglycan structure recognized. The relationship of fine structure and core protein structure to receptor binding is now known to be necessary for multiple cellular processes. Techniques used for genomics, proteomics, and cell biology have been successfully applied to the investigation of glycosaminoglycan signaling pathways. While some aspects of the structural requirements of glycosaminoglycans and proteoglycans for effective signaling have been identified, the reasons for why they participate in cellular processes and how the signaling pathways are initiated and regulated are still to be determined.

References:1. Silbert, J.E. and Sugumaran, G. Biosynthesis of chondroitin/dermatan sulfate. IUBMB

Life, 54, 177-86 (2002).2. Taylor, K.R., and Gallo, R.L., Glycosaminoglycans and their proteoglycans: host-

associated molecular patterns for initiation and modulation of inflammation. FASEB J., 20, 9-22 (2006).

3. Habuchi, H., et al., Sulfation pattern in glycosaminoglycan: does it have a code? Glycoconj. J., 21, 47-52 (2004).

4. Habuchi, H., et al., The occurrence of three isoforms of heparan sulfate 6-O-sulfotransferase having different specificities for hexuronic acid adjacent to the targeted N-sulfoglucosamine. J. Biol. Chem., 275, 2859-68 (2000).

5. Gallagher, J.T., Multiprotein signalling complexes: regional assembly on heparan sulphate. Biochem. Soc. Trans., 34, 438-41 (2006).

6. Lamanna, W.C., et al., The heparanome - the enigma of encoding and decoding heparan sulfate sulfation. J. Biotech., 129, 290-307 (2007).

7. Petitou, M., et al., 1976-1983, a critical period in the history of heparin: the discovery of the antithrombin binding site. Biochimie, 85, 83-89 (2005).

8. Nakato, H., and Kimata, K. Heparan sulfate fine structure and specificity of proteoglycan functions. Biochim. Biophys. Acta, 1573, 312-8 (2002).

9. Kato, M., et al., Physiological degradation converts the soluble syndecan-1 ectodomain from an inhibitor to a potent activator of FGF-2. Nat. Med., 4, 691-7 (1998).

10. Mundhenke, C., et al., Heparan sulfate proteoglycans as regulators of fibroblast growth factor-2 receptor binding in breast carcinomas. Am. J. Pathol., 160, 185-94 (2002).

11. Shipp, E.L. and Hseih-Wilson, L.C., Profiling the sulfation specificities of glycosaminoglycan interactions with growth factors and chemotactic proteins using microarrays. Chem. Biol., 14, 195-208 (2007).

12. Bournemann, D.J., et al., Abrogation of heparan sulfate synthesis in Drosophila disrupts the Wingless, Hedgehog and Decapentaplegic signaling pathways. Development, 131, 1927-1938 (2004).

13. Han, C., et al., Distinct and collaborative roles of Drosophila EXT family proteins in morphogen signalling and gradient formation. Development, 131, 1563-1575 (2004).

14. De Cat, B., et al., Processing by proprotein convertases is required for glypican-3 modulation of cell survival, Wnt signaling, and gastrulation movements. J. Cell Biol., 163, 625-635 (2003).

15. Halden, Y., et al., Interleukin-8 binds to syndecan-2 on human endothelial cells. Biochem. J., 377, 533-538 (2004).

16. Jones, M. et al., Heparan sulfate proteoglycan isoforms of the CD44 hyaluronan receptor induced in human inflammatory macrophages can function as paracrine regulators of fibroblast growth factor action. J. Biol. Chem., 275, 7964-74 (2000).

17. You, R.I., et al., Apoptosis of dendritic cells induced by decoy receptor 3 (DcR3). Blood, 111, 1480-8 (2008).

18. Narita, K., et al., HSulf-1 inhibits angiogenesis and tumorigenesis in vivo. Cancer Res., 66, 6025-32 (2006).

19. Sugaya, N., et al., 6-O-Sulfation of heparan sulfate differentially regulates various fibroblast growth factor-dependent signalings in culture. J. Biol. Chem., 283, 10366-76 (2008).

20. Ashikari-Hada, S., et al., Heparin regulates vascular endothelial growth factor165-dependent mitogenic activity, tube formation, and its receptor phosphorylation of human endothelial cells. Comparison of the effects of heparin and modified heparins. J. Biol. Chem., 280, 31508-15 (2005).

21. Nandini, C.D. and Sugahara, K., Role of the sulfation pattern of chondroitin sulfate in its biological activities and in the binding of growth factors. Adv. Pharmacol., 53, 253-79 (2006).

22. Sugahara, K., et al., Recent advances in the structural biology of chondroitin sulfate and dermatan sulfate. Curr. Opin. Struct. Biol., 13, 612-20 (2003).

23. Penc, S.F., et al., Dermatan sulfate released after injury is a potent promoter of fibroblast growth factor-2 function. J. Biol. Chem., 273, 28116-21 (1998).

24. Trowbridge, J.M., et al., Dermatan sulfate binds and potentiates activity of keratinocyte growth factor (FGF-7). J. Biol. Chem., 277, 42815-20 (2002).

25. Taylor, K.R., et al., Structural and sequence motifs in dermatan sulfate for promoting fibroblast growth factor-2 (FGF-2) and FGF-7 activity. J. Biol. Chem., 280, 5300-6 (2005).

26. Béchard, D., et al., Endocan is a novel chondroitin sulfate/dermatan sulfate proteoglycan that promotes hepatocyte growth factor/scatter factor mitogenic activity. J. Biol. Chem., 276, 48341-48349 (2001).

27. Catlow, K.R., et al., Interactions of hepatocyte growth factor/scatter factor with various glycosaminoglycans reveal an important interplay between the presence of iduronate and sulfate density. J. Biol. Chem., 283, 5235-5248 (2008).

28. Tiedemann, K., et al., Regulation of the chondroitin/dermatan fine structure by transforming growth factor-β1 through effects on polymer-modifying enzymes. Glycobiology, 15, 1277-1285 (2005).

29. Deepa, S.S., et al., Specific molecular interactions of oversulfated chondroitin sulfate E with various heparin-binding growth factors. J. Biol. Chem., 277, 43707-43716 (2002).

30. Purushothaman, A., et al., Functions of chondroitin sulfate/dermatan sulfate chains in brain development. Critical roles of E and iE disaccharide units recognized by a single chain antibody GD3G7. J. Biol. Chem., 282, 19442-19452 (2007).

31. Seidler, D.G. and Dreier, R., Decorin and its galactosaminoglycan chain: extracellular regulator of cellular function? IUBMB Life, 60, 729-33 (2008).

32. Bhattacharyya, S., et al., Distinct effects of N-acetylgalactosamine-4-sulfatase and galactose-6-sulfatase expression on chondroitin sulfates. J. Biol. Chem., 283, 9523-9530 (2008).

33. Hitchcock, A.M., et al., Comparative glycomics of connective tissue glycosaminoglycans. Proteomics, 8, 1384-1397 (2008).

34. Imberty, A., et al., Structural view of glycosaminoglycan-protein interactions. Carbohydr. Res., 342, 430-439 (2007).

35. Nairn, A.V., et al., Glycomics of proteoglycan biosynthesis in murine embryonic stem cell differentiation. J. Proteome Res., 6, 4374-87 (2007).

36. Kirkpatrick, C.A. and Selleck, S. B., Heparan sulfate proteoglycans at a glance. J. Cell Sci., 120, 1829-32 (2007).

http://www.cermav.cnrs.fr/glyco3d


Gly

cosa

min

og

lyca

n S

ulf

atio

n a

nd

Sig

nal

ing

Glycosaminoglycans and Proteoglycans

Glycosaminoglycans

Chondroitin sulfate B sodium saltDermatan sulfate sodium salt; β-Heparin [54328-33-5]

from porcine intestinal mucosa, ≥90%, lyophilized powder store at: 2-8°C

C3788-25MG 25 mg

C3788-100MG 100 mg

Chondroitin sulfate sodium salt from shark cartilage[9007-28-7]Natural polymer of β-glucuronic acid-(1→ 3)N-acetyl-β-galactosamine [β-GlcA-(1→3)-GalNAc-(1→4)]. Sulfation may occur at the 6-position and/or 4-position of the galactosamine moiety, with the potential of any individual disaccharide unit to be 6-sulfated, 4-sulfated, 4,6-disulfated, or unsulfated.

Ratio of β-GlcA-(1→3)-GalNAc-6-sulfate to β-GlcA-(1→3)-GalNAc-4-sulfate is determined by HPLC after enzymatic degradation. store at: 2-8°C

C4384-250MG 250 mg

C4384-1G 1 g

C4384-5G 5 g

C4384-25G 25 g

Chondroitin sulfate A sodium salt from bovine tracheaAlternating Co poly β-glucu ronic acid-(1→3)-N-acetyl-β-galacto s amine-4-sulfate-(1→4)[39455-18-0]

cell culture tested, lyophilized powderApprox. 70%; balance is chondroitin sulfate C

Appears to play a regulatory role for chondrocytes, neural cells, and some tumor cells

surface coverage .................................................................. 20-2000 μg/cm2

store at: 2-8°C

C9819-5G 5 g

C9819-25G 25 g

He para n sulfate sodium salt from bovine kidneyHeparitin sulfate sodium salt [57459-72-0]Constituent of membrane-associated proteoglycans suggested to contribute to cell-cell adhesion store at: 2-8°C

H7640-1MG 1 mg

H7640-5MG 5 mg

H7640-10MG 10 mg

He para n sulfate fast-moving fraction sodium salt from porcine intestinal mucosaHeparitin sulfate [57459-72-0]

≥90% (electrophoresis) store at: 2-8°C

H9902-1MG 1 mg

H9902-5MG 5 mg

q Heparin

Heparin ammonium salt from porcine intestinal mucosa[60800-63-7]

activity: ~140 USP units/mgH6279-25KU 25,000 units

H6279-100KU 100,000 units

H6279-250KU 250,000 units

H6279-500KU 500,000 units

H6279-1MU 1,000,000 units

Heparin lithium salt from porcine intestinal mucosa[9045-22-1]

activity: ≥150 USP units/mgH0878-100KU 100,000 units

H0878-1MU 1,000,000 units

Heparin sodium salt[9041-08-1]

activity: ≥140 USP units/mgLow calcium content

EDTA ......................................................................................none detectedCa .................................................................................................. ≤20 ppm store at: 2-8°C

H4784-250MG 250 mg

H4784-1G 1 g

Heparin sodium salt from bovine intestinal mucosa[9041-08-1]


H0777-50KU 50,000 units

H0777-100KU 100,000 units

H0777-250KU 250,000 units

H0777-500KU 500,000 units

H0777-1MU 1,000,000 units

Heparin sodium salt from porcine intestinal mucosa[9041-08-1]

Grade I-A, activity: ~170 USP units/mgH3393-10KU 10,000 units

H3393-25KU 25,000 units

H3393-50KU 50,000 units

H3393-100KU 100,000 units

H3393-250KU 250,000 units

H3393-500KU 500,000 units

H3393-1MU 1,000,000 units


H9399-50KU 50,000 units

H9399-100KU 100,000 units

H9399-250KU 250,000 units

H9399-500KU 500,000 units

H9399-1MU 1,000,000 units

H9399-5MU 5,000,000 units

Heparin sodium salt from porcine intestinal mucosa[9041-08-1]

activity: ≥160 IU/mgCrude

Unbleached

H5515-25KU 25,000 units

H5515-100KU 100,000 units

H5515-250KU 250,000 units



http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=C3788&Brand=SIGMA








http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=H7640&Brand=SIGMA












http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=H4784&Brand=SIAL


























Glyco

samin

og

lycan Su

lfation

and

Sign

aling

Heparin sodium salt from porcine intestinal mucosa[9041-08-1]Low molecular weight

average mol wt ~3,000Depolymerized by peroxidolysis (free-radical induced cleavage).

activity: <60 units/mg

H3400-50MG 50 mg

H3400-100MG 100 mg

H3400-250MG 250 mg

H3400-1G 1 g

mol wt 4,000-6,000 DaH8537-50MG 50 mg

H8537-100MG 100 mg

H8537-250MG 250 mg

H8537-1G 1 g

Heparin p

Hyaluronan bio tin sodium salt

Hyaluronic acid bio tin

From cocks comb or fermentation.

Highly sensitive probe for detecting nanogram levels of hyaluronan binding proteins (hyaladherins).

mol wt ~700 kDa

>97%, soluble powder store at: 2-8°C

B1557-5MG 5 mg

q Hyaluronic acidPoly(β-glucu ronic acid-[1→3]-β-N-acetyl gluco s amine-[1→4]), alternating [9067-32-7]

Hyaluronic acid sodium salt from bovine vitreous humor(C14H21NaNO11)n

High molecular weight polymer composed of repeating dimeric units of glucuronic acid and N-acetyl glucosamine which forms the core of complex proteoglycan aggregates found in extracellular matrix. store at: −20°C

H7630-10MG 10 mg

H7630-50MG 50 mg

Hyaluronic acid sodium salt from rooster comb store at: −20°C

H5388-100MG 100 mg

H5388-250MG 250 mg

H5388-1G 1 g

Hyaluronic acid sodium salt from Streptococcus equi

BioChemika protein ...................................................................................................≤1% store at: −20°C

53747-1G 1 g

53747-10G 10 g

Hyaluronic acid p

Proteoglycans

Aggrecan from bovine articular cartilage

lyophilized powderMajor structural proteoglycan of cartilage extracellular matrix. Large proteoglycan with a molecular weight greater than 2,500 kDa. Approximately 100-150 glycosaminoglycan (GAG) chains are attached to the core protein (210-250 kDa). The majority of the GAG chains are chondroitin/dermatan sulfate with the remainder being keratan sulfate. This structural molecule produces a rigid, reversibly deformable gel that resists compression. It combines with hyaluronic acid to form very large macromolecular complexes. Addition of small amounts (0.1-2% w/w) of hyaluronic acid to an aggrecan solution (2mg/ml) results in the formation of a complex with an increased hydrodynamic volume and in a significant increase (30-40%) in the relative viscosity of the solution. Aggrecan is a critical component for cartilage structure and the function of joints. The synthesis and degradation of aggrecan are being investigated for their roles in cartilage deterioration during joint injury, disease, and aging. Contains three globular domains, G1, G2, and G3, that are involved in aggregation and hyaluronan binding, cell adhesion, and chondrocyte apoptosis.

Associated gene(s): AGC1 (280985)

Proteoglycans are extracted from articular cartilage with guanidine hydrochloride. Further purification includes gel filtration and ion exchange chromatography. The product is dialyzed against water and sterile-filtered prior to lyophilization.

salt .........................................................................................essentially free store at: −20°C

A1960-1MG 1 mg

Biglycan from bovine articular cartilage

Interacts with collagen type I, as well as with fibronectin and TGF-β.

essentially salt-free, lyophilized powder ( from a sterile-filtered solution)

Associated gene(s): BGN (280733)

mol wt 200-350 kDa (proteoglycan consisting of a 45 kDa core protein and two chrondroitin/dermatan sulfate glycosaminoglycan chains) store at: −20°C

B8041-.5MG 0.5 mg

Decorin from bovine articular cartilage

salt-free, lyophilized powderDecorin interacts with collagen type I and II, fibronectin, thrombospondin and TGF-β.

Decorin is an approx. 100 kDa proteoglycan consisting of a 40 kDa core protein and one chondroitin or dermatan sulfate glycosaminoglycan chain.

Associated gene(s): DCN (280760) store at: −20°C

D8428-.5MG 0.5 mg















http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=53747&Brand=FLUKA


http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=A1960&Brand=SIGMA

http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=B8041&Brand=SIGMA

http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=D8428&Brand=SIGMA


Gly

cosa

min

og

lyca

n S

ulf

atio

n a

nd

Sig

nal

ing

Glypican 3 8

Sulfate proteoglycan attached to the cell surface by a glycosylphosphatidylinositol (GPI) anchor. Regulates cell survival and inhibits cell proliferation during normal development.1 Potential use as marker for early disease detection.2

Lit cited: 1. Song, H.H. et al., The loss of glypican-3 induces alterations in Wnt signaling. J. Biol. Chem. 280, 2116-2125 (2005); 2. Wang, X.Y. et al., Glypican-3 expression in hepatocellular tumors: diagnostic value for preneoplastic lesions and hepatocellular carcinomas. Hum. Pathol. 37, 1435-1441 (2006);

Glypican 3, Recombinant human

Under non-reducing conditions in SDS-PAGE, the glycanated glypican 3 appears as a smear with an apparent molecular mass of 60 - 100 kDa.

Lyophilized from a 0.2 μm filtered solution in PBS containing 50 μg of bovine serum albumin per 1 μg of cytokine.

recombinant, expressed in mouse NSO cells store at: −20°C

G1921-50UG 50 μg

He para n sulfate proteoglycan

HSPG

Extracellular matrix component that binds to fibroblast growth factors, vascular endothelial growth factor (VEGF) and VEGF receptors through its sugar moiety. Acts as a docking molecule for matrilysin (MMP-7) and other matrix metalloproteinases.

≥400 μg/mL glycosaminoglycanComposed of a core protein covalently bound to heparan sulfate chains.

For cell culture use.

Isolated from basement membrane of Engelbreth-Holm-Swarm mouse sarcoma.

Solution in 50 mM Tris HCl, 150 mM NaCl, 1 mM EDTA, 0.1 mM PMSF, pH 7.4, containing ≥400 μg protein per ml.

Uronic acid ................................................................................ ≥100 μg/mLship: dry ice store at: −20°C

H4777-.1MG 0.1 mg

q ProteoglycanChromatographically purified using 7M urea on DEAE-cellulose by modified procedure by Antonopoulos.1

Lit cited: 1. Antonopoulos, C.A., et al., Biochim. Biophys. Acta 338, 108-119 (1974);

Proteoglycan from bovine nasal septum store at: −20°C

P5864-10MG 10 mg

Proteoglycan from chicken sternal cartilage

solid store at: −20°C

P5989-2MG 2 mg

Proteoglycan p

Tissue Inhibitor of Met allo protein ase-1 human

TIMP-1

TIMP-1 has greater binding efficiency to MMP-9, MMP-1, and MMP-3 than the other MMPs.

Associated gene(s): TIMP1 (7076)

recombinant, expressed in CHO cells, ~500 μg/mL protein, buffered aqueous solution

Supplied in 0.01 M sodium phosphate buffer pH 7.3, 0.15 M NaCl.

mol wt ~29 kDaship: wet ice store at: −20°C

T8947-5UG 5 μg

Trans form ing Growth Factor-β Soluble Receptor III human

TGF-β sRIII

>97% (SDS-PAGE), recombinant, expressed in mouse NSO cells, lyophilized powder

Lyophilized from a 0.2 μm filtered solution in phosphate buffered saline containing 5 mg bovine serum albumin

The receptor-mediated activity is measured by its ability to inhibit the TGF-β2 bioactivity in HT-2 cells.

Endotoxin ............................................................................................tested store at: −20°C

T4567-.1MG 0.1 mg



http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=G1921&Brand=SIGMA


http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=P5864&Brand=SIGMA


http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=T8947&Brand=SIGMA

http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=T4567&Brand=SIGMA


Glyco

samin

og

lycan Su

lfation

and

Sign

aling

Glycosaminoglycan Degrading Enzymes

Chondroitinases

Name Specific Activity Unit Definition Cat. No.

Chondroitinase ABC from Proteus vulgaris 0.3-3 units/mg solid using chondroitin sulfate C as substrate

One unit will liberate 1.0 μmole of 2-acetamido-2-deoxy-3-O-(β-D-gluc-4-ene-pyranosyluronic acid)-4-O-sulfo-D-galactose from chondroitin sulfate A or 1.0 μmole of 2-acetamido-2-deoxy-3-O-(β-D-gluc-4-ene-pyranosyluronic acid)-6-O-sulfo-D-galactose from chondroitin sulfate C per min at pH 8.0 at 37 °C.

C2905-2UNC2905-5UNC2905-10UN

Lyophilized powder containing Tris buffer salts

Chondroitinase ABC from Proteus vulgaris 50-250 units/mg protein using chondroitin sulfate C as substrate

One unit will liberate 1.0 μmole of 2-acetamido-2-deoxy-3-O-(β-D-gluc-4-ene-pyranosyluronic acid)-4-O-sulfo-D-galactose from chondroitin sulfate A or 1.0 μmole of 2-acetamido-2-deoxy-3-O-(β-D-gluc-4-ene-pyranosyluronic acid)-6-O-sulfo-D-galactose from chondroitin sulfate C per min at pH 8.0 at 37 °C.

C3667-5UNC3667-10UN

lyophilized powder

Chondroitinase AC from Flavobacterium heparinum 0.5-1.5 units/mg solid using chondroitin sulfate A as substrate, also cleaves chondroitin sulfate C

One unit will cause a ΔA232 of 1.0 per minute due to the release of unsaturated disaccharide from chondroitin sulfate A at pH 7.3 at 37 °C. Reaction volume: 3.1 ml (light path 1 cm).

C2780-5UN

lyophilized powder

Chondroitinase B from Flavobacterium heparinum 100-300 units/mg solid One unit will form 0.1 μmole of unsaturated uronic acid per hr at pH 7.5 at 25°C using chondroitin sulfate B as substrate.

C8058-50UN

lyophilized powder (with BSA as stabilizer)

Chondroitinase C from Flavobacterium heparinum ≥200 units/mg solid One unit will form 0.1 μmole of unsaturated uronic acid per hr at pH 8.0 at 25 °C using chondroitin sulfate C as substrate.

C0954-75UN

lyophilized powder

Heparinases

Name Specific Activity Unit Definition Cat. No.

Heparinase I from Flavobacterium heparinum 200-600 unit/mg solid One unit will form 0.1 μmole of unsaturated uronic acid per hr at pH 7.5 at 25 °C. One International Unit (I.U.) is equivalent to approx. 600 Sigma units.

H2519-50UNH2519-100UNH2519-250UN

Heparinase II from Flavobacterium heparinum 100-300 units/mg solid One unit will form 0.1 μmole of unsaturated uronic acid per hr at pH 7.0 at 25 °C. One International Unit (I.U.) is equivalent to approx. 600 Sigma units.

H6512-10UNH6512-25UNH6512-100UN

lyophilized powder

Heparinase III from Flavobacterium heparinum 200-600 unit/mg solid One unit will form 0.1 μmole of unsaturated uronic acid per hr at pH 7.5 at 25 °C. One International Unit (I.U.) is equivalent to approx. 600 Sigma units.

H8891-5UNH8891-10UNH8891-50UN

Hyaluronidases

Name Specific Activity Analysis Cat. No.

Hyaluronidase from bovine testes, Type I-S 400-1000 units/mg solid One unit is based on the change in absorbance at 600 nm (change in turbidity) of a USP reference standard hyaluronidase which is assayed concurrently with each lot of this product.

H3506-100MGH3506-500MGH3506-1GH3506-5G

Hyaluronidase from bovine testes, Type IV-S 750-3000 units/mg solid One unit will cause a change in A600 of 0.330 per minute at pH 5.7 at 37°C H3884-50MGH3884-100MGH3884-500MGH3884-1G

Hyaluronidase from bovine testes, Type VIII ~300 units/mg One unit will cause a change in A600 nm of 0.330 per minute at pH 5.7 at 37 °C H3757-100MG

Hyaluronidase from bovine testes, Type VI-S 3,000-15,000 units/mg solid One unit is based on the change in absorbance at 600 nm (change in turbidity) of a USP reference standard hyaluronidase which is assayed concurrently with each lot of this product.

H3631-3KUH3631-15KUH3631-30KU

Hyaluronidase from sheep testes, Type V ≥1,500 units/mg solid One unit will cause a change in A600 nm of 0.330 per minute at pH 5.7 at 37 °C H6254-500MGH6254-1G

Hyaluronidase from sheep testes, Type II ≥300 units/mg One unit will cause a change in A600 nm of 0.330 per minute at pH 5.7 at 37 °C H2126-100MGH2126-500MGH2126-1GH2126-5G

Lyophilized powder containing lactose

Hyaluronidase from Streptomyces hyalurolyticus - - H1136-1AMP






































Gly

cosa

min

og

lyca

n S

ulf

atio

n a

nd

Sig

nal

ing

Prestige Antibodies

Product Name Host Form Gene Symbol Species Reactivity Application Cat. No.

Anti-BGN 8 rabbit affinity isolated antibody BGN, human human IHC (p)PA

HPA003157-100UL

Anti-CHST2 8 rabbit affinity isolated antibody CHST2, human human IHC (p)PAWB

HPA013313-100UL

Anti-CSPG4 8 rabbit affinity isolated antibody CSPG4, human human IHC (p)PA

HPA002951-100UL

Anti-DCN 8 rabbit affinity isolated antibody DCN, human human IHC (p)PA

HPA003315-100UL

Anti-DSE 8 rabbit affinity isolated antibody DSE, human human IHC (p)PA

HPA014764-100UL

Anti-FGF1 8 rabbit affinity isolated antibody FGF1, human human IHC (p)PA

HPA003265-100UL

Anti-FGF4 8 rabbit affinity isolated antibody FGF4, human human IHC (p)PA

HPA011209-100UL

Anti-GPC6 8 rabbit affinity isolated antibody GPC6, human human IHC (p)PAWB

HPA017671-100UL

Anti-HS3ST1 8 rabbit affinity isolated antibody HS3ST1, human human IHC (p)PA

HPA002237-100UL

Anti-HSPG2 8 rabbit affinity isolated antibody HSPG2, human human IHC (p)PA

HPA018892-100UL

Anti-SDC1 8 rabbit affinity isolated antibody SDC1, human human IHC (p)PA

HPA006185-100UL


HPA017087-100UL


HPA005716-100UL

Anti-SRGN 8 rabbit affinity isolated antibody SRGN, human human IHC (p)PA

HPA000759-100UL

Anti-SULF2 8 rabbit affinity isolated antibody SULF2, human human IHC (p)PA

HPA002325-100UL

Immunohistochemistry

Anti-BGN: Cat. No. HPA003157: Immunoperoxidase staining of formalin-fixed, paraffin-embedded human soft tissue showing positive staining of chondrocytes in the cartilage.


Anti-HS3ST1: Cat. No. HPA002237: Immunoperoxidase staining of formalin-fixed, paraffin-embedded human cerebral cortex tissue showing positive staining of neuronal and non-neuronal cells.


Anti-SULF2: Cat. No. HPA002325: Immunoperoxidase staining of formalin-fixed, paraffin-embedded human heart muscle tissue showing strong cytoplasmic staining of myocytes.

Antibodies

Prestige Antibodies™

Prestige Antibodies Powered by Atlas Antibodies are developed and validated by the Human Proteome Resource (HPR) project (proteinatlas.org). Each antibody is tested by immunohistochemistry against hundreds of normal and disease tissues. These images can be viewed on the Human Protein Atlas (HPA) site.

For each antibody, the site includes multiple immunohistochemical images and a summary of the protein expression and staining level found using different cell lines and tissues. In certain tumor groups, subtypes have been included and efforts have been made to include high and low grade malignancies where applicable. Tumor heterogenity and inter-individual differences are reflected in diverse expression of proteins resulting in variable immunohistochemical staining patterns. The antibodies are also tested using protein array and Western blotting.

For application protocols and other useful information about Prestige Antibodies and the HPA, visit sigma.com/prestige.



http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=HPA003157&Brand=SIGMA


















http://www.sigma.com/prestige


Glyco

samin

og

lycan Su

lfation

and

Sign

aling

Antibodies to PG Core Proteins

Product Name Host Clone Form Application Cat. No.

Monoclonal Anti-AGC1 mouse 2A8 purified immunoglobulin ELISA (i)WB

WH0000176M1-100UG

Monoclonal Anti-DCN mouse 3H4-1F4 purified immunoglobulin ELISA (i)IHC (p)

WB

WH0001634M1-100UG

Monoclonal Anti-GPC5 mouse 1C9 purified immunoglobulin ELISA (i) WH0002262M1-100UG

Antibodies to GAG Synthesis/Modification Enzymes


Monoclonal Anti-ARSB mouse 1A4 purified immunoglobulin ELISA (i)WB

WH0000411M2-100UG

Monoclonal Anti-EXT1 mouse 5A5 purified immunoglobulin ELISA (i)WB

WH0002131M1-100UG

Monoclonal Anti-EXT2 mouse 3G6 purified immunoglobulin ELISA (i)WB

WH0002132M1-100UG

Monoclonal Anti-NDST1 mouse 1G10 purified immunoglobulin ELISA (i)IHC (p)

WB

WH0003340M1-100UG

Monoclonal Anti-NDST3 mouse 5B9 purified immunoglobulin ELISA (i)WB

WH0009348M1-100UG

Monoclonal Anti-CHST3 mouse 1C4 purified immunoglobulin ELISA (i)WB

WH0009469M1-100UG

Monoclonal Anti-SART2 mouse 6D4 purified immunoglobulin ELISA (i)WB

WH0029940M3-100UG

Monoclonal Anti-CHST11 mouse 4F1 purified immunoglobulin ELISA (i)WB

WH0050515M1-100UG

Additional Antibodies


Monoclonal Anti-Chondroitin Sulfate mouse CS-56 ascites fluid ARRIF (i)

C8035-.2MLC8035-.5ML

Application Abbreviation Table

Application Abbreviation

ANA-indirect immunofluorescence IF (ANA)

Capture ELISA ELISA (c)

Direct ELISA ELISA (d)

Direct immunofluorescence IF (d)

Dot blot DB

Dot immunobinding DIBA

Electron microscopy EM

Enzyme immunoassay EIA

Flow cytometry FACS

Immunoblotting WB

Immunoblotting (chemiluminescent) WB CL

Immunocytochemistry ICC

Immunoelectrophoresis IEP

Application Abbreviation

Immunohistochemistry IHC

Immunohistochemistry (formalin-fixed, paraffin-embedded sections) IHC (p)

Immunohistochemistry (frozen sections) IHC (f)

Immunoprecipitation IP

Indirect ELISA ELISA (i)

Indirect immunofluorescence IF (i)

Microarray ARR

Neutralization Neutral

Ouchterlony double diffusion ODD

Particle immunofluorescence PIFA

Quantitative precipitin assay QPA

Radioimmunoassay RIA

http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=WH0000176M1&Brand=SIGMA














Gly

cosa

min

og

lyca

n S

ulf

atio

n a

nd

Sig

nal

ing

Glycosaminoglycan DisaccharidesIn the following listings, ΔUA = 4-deoxy-L-threo-hex-4-enopyranosyluronic acid; GalNAc = N-acetyl-D-galactosamine; GlcN = D-glucosamine; GlcNAc = N-acetyl-D-glucosamine; NS = N-sulfo-; 2S = 2-sulfate; 4S = 4-sulfate; 6S = 6-sulfate. The uronic acid moiety loses its chirality on cleavage from the native polysaccharide.

Name Synonyms Formula Formula Weight Cat. No.

Chondroitin disaccharide Δdi-0S sodium salt α-ΔUA-[1→3]-GalNAc C14H20NNaO11 401.30 C3920-5MGC3920-10MG

Chondroitin disaccharide Δdi-4S sodium salt α-ΔUA-[1→3]-GalNAc-4S C14H19NNa2O14S 503.34 C4045-5MGC4045-10MG

Chondroitin disaccharide Δdi-6S sodium salt α-ΔUA-[1→3]-GalNAc-6S C14H19NNa2O14S 503.34 C4170-5MGC4170-25MG

Chondroitin disaccharide Δdi-UA-2S sodium salt α-ΔUA-2S-[1→3]-GalNAc C14H19NNa2O14S 503.34 C5820-1MG

Heparin disaccharide I-A sodium salt α-ΔUA-2S-[1→4]-GlcNAc-6S C14H18NNa3O17S2 605.39 H9517-1MG

Heparin disaccharide I-H sodium salt α-ΔUA-2S-[1→4]-GlcN-6S C12H17NNa2O16S2 541.37 H8892-1MG

Heparin disaccharide I-S sodium salt α-ΔUA-2S-[1→4]-GlcNS-6S C12H15NNa4O19S3 665.40 H9267-1MGH9267-5MG

Heparin disaccharide II-H sodium salt α-ΔUA-[1→4]-GlcN-6S C12H18NNaO13S 439.33 H9017-1MG

Heparin disaccharide II-S sodium salt α-ΔUA-[1→4]-GlcNS-6S C12H16NNa3O16S2 563.35 H1020-.5MG

Heparin disaccharide III-H sodium salt α-ΔUA-2S-[1→4]-GlcN C12H18NNaO13S 439.33 H9142-1MG

Heparin disaccharide III-S sodium salt α-ΔUA-2S-[1→4]-GlcNS C12H16NNa3O16S2 563.35 H9392-1MG

Heparin disaccharide IV-A sodium salt α-ΔUA-[1→4]-GlcNAc C14H20NNaO11 401.30 H0895-.5MG

Heparin disaccharide IV-H α-ΔUA-[1→4]-GlcN C12H19NO10 337.28 H9276-1MG

Monosaccharide Sulfates

Name Synonyms Formula Formula Weight Cat. No.

N-Acetyl-D-galactosamine 6-sulfate sodium salt GalNAc-6S C8H14NNaO9S 323.25 51947-1MG

N-Acetyl-D-glucosamine 6-sulfate sodium salt GlcNAc-6S C8H14NNaO9S 323.25 44001-25MG

D-Galactosamine 2-sulfate sodium salt 2S-GalN C6H12NNaO8S 281.22 12662-5MG

D-Galactose 4-sulfate sodium salt Gal-4S C6H11NaO9S 282.20 90572-1MG

D-Galactose 6-sulfate sodium salt Gal-6S C6H11NaO9S 282.20 05801-1MG

D-Glucosamine 2-sulfate sodium salt GlcN-2S C6H12NNaO8S 281.22 G7889-100MG

D-Glucosamine 3-sulfate GlcN-3S C6H13NO8S 259.23 11631-25MG

D-Glucosamine 6-sulfate GlcN-6S C6H13NO8S 259.23 G8641-25MG

D-Glucose 3-sulfate sodium salt Glc-3S C6H11NaO9S 282.20 G9909-10MGG9909-50MG

D-Mannose 6-sulfate sodium salt Man-6S C6H11NaO9S 282.20 14652-50MG




















http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=51947&Brand=SIGMA












Dextran

and

Related

Polysacch

arides

DextranHistorically, dextrans had been long recognized as contaminants in sugar processing and other food production. The formation of dextran in wine was shown by Pasteur to be due to the activity of microbes.1 The name dextran was created by Scheibler in 1874, who demonstrated dextran was a carbohydrate with the formula (C6H10O6)n and a positive optical rotation.2

Dextrans are polysaccharides with molecular weights ≥1,000 Dalton, which have a linear backbone of α-linked D-glucopyranosyl repeating units. Three classes of dextrans can be differentiated by their structural features. The pyranose ring structure contains five carbon atoms and one oxygen atom. Class 1 dextrans contain the α(1→6)-linked D-glucopyranosyl backbone modified with small side chains of D-glucose branches with α(1→2), α(1→3), and α(1→4)-linkage (see Figure 1). The class 1 dextrans vary in their molecular weight, spatial arrangement, type and degree of branching, and length of branch chains,3-5 depending on the microbial producing strains and cultivation conditions.6,7 Isomaltose and isomaltotriose are oligosaccharides with the class 1 dextran backbone structure. Class 2 dextrans (alternans) contain a backbone structure of alternating α(1→3) and α(1→6)-linked D-glucopyranosyl units with α(1→3)-linked branches. Class 3 dextrans (mutans) have a backbone structure of consecutive α(1→3)-linked D-glucopyranosyl units with α(1→6)-linked branches. One and two-dimensional NMR spectroscopy techniques have been utilized for the structural analysis of dextrans.8

The secretion of dextrans provides an opportunity for bacteria to modulate adhesion, e.g. in tooth decay, by having a softer or more rigid bacterial cell surface, depending on the polysaccharide itself and the pH and ionic strength. Low bacterial adhesion occurs at low salt conditions with more rigid polysaccharides and a softer surface, while high bacterial adhesion is obtained with more flexible polysaccharides and a rigid bacterial surface. Polymer elasticity is important for structural integrity. and the pyranose ring is the structural unit controlling the elasticity of the polysaccharide. This elasticity results from a force-induced elongation of the ring structure and a final transition from a chair-like to a boat-like conformation of the glucopyranose ring, which plays an important role in accommodating mechanical stress and modulating ligand binding in biological systems.9 Laboratory experiments have demonstrated that cleavage of the pyranose rings of dextran, amylose, and pullulan convert these different polysaccharide chains into similar structures where all the bonds of the polymer backbone can rotate and align under force. After ring cleavage, single molecules of dextran, amylose, and pullulan display identical elastic behavior as measured by atomic force microscopy.

OHO

O

HOOH

O

OHOHO

OH

O

OHOHO

OH

O

OHOHO

OH

O

OHOHO

OH

O

a-1,4-linkedD-Glucopyranosideside chains

a-1,3-linkedD-Glucopyranosideside chains

a-1,2-linkedD-Glucopyranosideside chains O

HOOH

O

HO

1

1

1

1

1

1

2

2

2

2

2

2

3

3

3

3

3

3

4

4

4

4

4

4

5

5

5

5

5

5

6

6

6

6

6

6

Figure 1. General structure of class 1 dextrans consisting of a linear backbone of α(1→6)-linked D-glucopyranosyl repeating units. The dextran may have branches of smaller chains of D-glucose linked to the backbone by α(1→2)- , α(1→3)- or α(1→4)- glycosidic bonds.

Dextrans are found as bacterial extracellular polysaccharides. They are synthesized from sucrose by beneficial lactic acid bacteria, such as Leuconostoc mesenteroides and Lactobacillus brevis, but also by the dental plaque-forming species Streptococcus mutans. Bacteria employ dextran in biofilm formation10 or as protective coatings, e.g., to evade host phagocytes in the case of pathogenic bacteria.11

The physical and chemical properties of purified dextrans vary depending on the microbial strains from which they are produced and by the production method, but all are white and tasteless solids. Dextrans have high water solubility and the solutions behave as Newtonian fluids. Solution viscosity depends on concentration, temperature, and molecular weight, which have a characteristic distribution.

Dextran and Related Polysaccharides


Dex

tran

an

d R

elat

ed P

oly

sacc

har

ides

The long history of the safety of dextrans has allowed them to be used as additives to food and chemicals, and in pharmaceutical and cosmetics manufacturing.12 Dextrans have been investigated for the targeted and sustained delivery of drugs, proteins, enzymes, and imaging agents.13 In medicine, clinical grades of dextrans with a molecular weight range of 75-100 kDa have been used as blood-plasma volume expanders in transfusions.14 Other applications include the use of dextrans with polyethylene glycol as components of aqueous two-phase systems for the extraction of biochemicals. The hydroxyl groups present in dextran offer many sites for derivatization, and these functionalized glycoconjugates represent a largely unexplored class of biocompatible and environmentally safe compounds.

Cross-linked dextran beads are widely used for chromatography in biochemical research and industry. The classic application of cross-linked dextrans is as gel filtration media in packed-bed columns for the separation and purification of biomolecules with molecular weights in the range of 0.7-200 kDa.15-17 Ion exchange chromatography utilizes dextran that has been derivatized with positively or negatively charged moieties such as carboxymethyl (CM), diethylaminoethyl (DEAE), diethyl(2-hydroxy propyl) amino ethyl (QAE), and sulfopropyl (SP).

Sigma® offers a large variety of dextrans with high polydispersity and dextran molecular weight standards with low polydispersity (Mw/Mn values close to 1.0).

Other PolysaccharidesPullulans are structural polysaccharides primarily produced from starch by the fungus Aureobasidium pullulans.18,19 Pullulans are composed of repeating α(1→6)-linked maltotriose (D-glucopyranosyl-α(1→4)-D-glucopyranosyl-α(1→4)-D-glucose) units with the inclusion of occasional maltotetraose units.20 Diffusion-ordered NMR spectroscopy has been used to achieve a simple estimation of the molecular weight of pullulan.21 The solution properties of pullulan in water have been studied, and it was confirmed that pullulan molecules behave as random coils in aqueous solution.22

Dextrins are composed of D-glucopyranosyl units but have shorter chain lengths than dextrans. They start with a single α(1→6) bond, but continue linearly with α(1→4)-linked D-glucopyranosyl units. Dextrins are usually mixtures derived from the hydrolysis of starch and have found widespread use in the food, paper, textile, and pharmaceutical industries.

Dextran sulfates are derived from dextran via sulfation. They have become indispensable components in many molecular biology techniques, including the transfer of large DNA fragments from agarose gels and rapid hybridization,23 precipitation procedures for the quantitation of high-density lipoprotein cholesterol,24 and inhibition of virion binding to CD4+ cells.25

References:1. Pasteur, L., Bull. Soc. Chim. Paris, 30-31 (1861). 2. Scheibler, C., Z. Ver. Dtsch. Zucker-Ind., 24, 309-335 (1874).3. Robyt, J.F., in: Encyclopedia of Polymer Sci. Eng., J.I.Kroschwitz (ed.), 4, 752-767

(1986), Wiley-VCH. 4. Cheetham, N.W.H., et al., Dextran structural details from high-field proton NMR

spectroscopy. Carbohydr. Polym. 14, 149-158 (1990).5. Naessens, M., et al., Leuconostoc dextransucrase and dextran: production, properties

and applications. J. Chem. Technol. Biotechnol., 80, 845-860 (2005).6. Kim, D., et al., Dextran molecular size and degree of branching as a function of su-

crose concentration, pH, and temperature of reaction of Leuconostoc mesenteroides B-512FMCM dextransucrase. Carbohydr. Res., 338, 1183-11889 (2003).

7. Côté, G.L., and Leathers, T.D., A method for surveying and classifying Leuconostoc spp. glucansucrases according to strain-dependent acceptor product patterns. J. Ind. Microbiol. Biotechnol. 32, 53-60 (2005).

8. Maina, N.H., et al., NMR spectroscopic analysis of exopolysaccharides produced by Leuconostoc citreum and Weissella confusa. Carbohydr. Res., 343, 1446-1455 (2008).

9. Marszalek, P.E., et al., Polysaccharide elasticity governed by chair-boat transitions of the glucopyranose ring. Nature, 396, 661-664 (1998).

10. Banas, J.A., and Vickermann, M.M., Glucan-binding proteins of the oral streptococci. Crit. Rev. Oral Biol. Med., 14, 89-99 (2003).

11. Meddens, M.J., et al., Br. J. Exp. Pathol., 65, 257-265 (1984).12. Kato, I., Fragrance J., 33, 59-64 (2005).13. Mehvar, R., Dextrans for targeted and sustained delivery of therapeutic and imaging

agents. J. Controlled Release, 69, 1-25 (2000).14. Terg, R., et al., Pharmacokinetics of Dextran-70 in patients with cirrhosis and ascites

undergoing therapeutic paracentesis. J. Hepatol., 25, 329-333 (1996).15. Porsch, B., and Sundelöf, L.-O., Size-exclusion chromatography and dynamic light

scattering of dextrans in water: Explanation of ion-exclusion behaviour. J. Chro-matogr. A, 669, 21-30 (1994).

16. Neyestani, T.R., et al., Isolation of α-lactalbumin, β-lactoglobulin, and bovine serum albumin from cow’s milk using gel filtration and anion-exchange chromatography including evaluation of their antigenicity. Protein Expres. Purif., 29, 202-208 (2003).

17. Penzol, G., et al., Use of dextrans as long and hydrophilic spacer arms to improve the performance of immobilized proteins acting on macromolecules. Biotechnol. Bioeng., 60, 518-523 (1998).

18. Gibson, L.H., and Coughlin, R.W., Optimization of high molecular weight pullulan production by Aureobasidium pullulans in batch fermentations. Biotechnol. Prog., 18, 675-678 (2002).

19. Leathers, T.D., Biotechnological production and applications of pullulan. Appl. Micro-biol. Biotechnol., 62, 468-473 (2003).

20. Catley, B.J., Pullulan, a relationship between molecular weight and fine structure. FEBS Lett., 10, 190-193 (1970).

21. Viel, S., et al., Diffusion-ordered NMR spectroscopy: a versatile tool for the molecular weight determination of uncharged polysaccharides. Biomacromolecules, 4, 1843-1847 (2003).

22. Nishinari, K., et al., Solution properties of pullulan. Macromolecules, 24, 5590-5593 (1991).

23. Wahl, G.M., et al., Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc. Natl. Acad. Sci. USA, 76, 3683-3687 (1979).

24. Warnick, G.R., et al., Dextran sulfate-Mg2+ precipitation procedure for quantitation of high-density-lipoprotein cholesterol. Clin. Chem., 28, 1379-1388 (1982).

25. Mitsuya H., et al., Dextran sulfate suppression of viruses in the HIV family: inhibition of virion binding to CD4+ cells. Science, 240, 646-649 (1988).




Dextran

and

Related

Polysacch

arides

DextransDextranFor General Applications

Name Mol Wt Cat. No.

Dextran Mr ~1,500 31394-5G31394-25G31394-100G

BioChemikaenzymatic synth.

Dextran from Leuconostoc spp. Mr ~6,000 31388-25G31388-100G31388-500G

BioChemika

Dextran from Leuconostoc spp. Mr 15,000-25,000 31387-25G31387-100G31387-500G

BioChemika


BioChemika


BioChemika

Dextran from Leuconostoc spp. Mr ~100,000 09184-10G-F09184-50G-F09184-250G-F

BioChemika

2nd edition

Tools for Glycoproteomics and Glycomics

Glycan Labeling and Analysis

Glycoprotein Purification and Detection

Chemical and Enzymatic Deglycosylation

Enzymatic Synthesis and Degradation

Glycobiology Analysis Manual

JFY02060-4013200127

World Headquarters3050 Spruce St., St. Louis, MO 63103(314) 771-5765sigma-aldrich.com

Order/Customer Service (800) 325-3010 • Fax (800) 325-5052

Technical Service (800) 325-5832 • sigma-aldrich.com/techservice

Development/Bulk Manufacturing Inquiries (800) 244-1173

©2007 Sigma-Aldrich Co. All rights reserved. SIGMA, , SAFC, , SIGMA-ALDRICH, ALDRICH, , FLUKA, , and SUPELCO, are trademarks belonging to Sigma-Aldrich Co. and its affiliate Sigma-Aldrich Biotechnology, L.P. Sigma brand products are sold through Sigma-Aldrich, Inc. Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip.

ArgentinaSIGMA-ALDRICH DE ARGENTINA S.A.Free Tel: 0810 888 7446Tel: (+54) 11 4556 1472Fax: (+54) 11 4552 1698

AustraliaSIGMA-ALDRICH PTY LTD.Free Tel: 1800 800 097 Free Fax: 1800 800 096Tel: (+61) 2 9841 0555Fax: (+61) 2 9841 0500

AustriaSIGMA-ALDRICH HANDELS GmbHTel: (+43) 1 605 81 10Fax: (+43) 1 605 81 20

BelgiumSIGMA-ALDRICH NV/SA.Free Tel: 0800 14747Free Fax: 0800 14745Tel: (+32) 3 899 13 01Fax: (+32) 3 899 13 11

BrazilSIGMA-ALDRICH BRASIL LTDA.Free Tel: 0800 701 7425Tel: (+55) 11 3732 3100Fax: (+55) 11 5522 9895

CanadaSIGMA-ALDRICH CANADA LTD.Free Tel: 1800 565 1400Free Fax: 1800 265 3858Tel: (+1) 905 829 9500Fax: (+1) 905 829 9292

ChinaSIGMA-ALDRICH (SHANGHAI) TRADING CO. LTD.Free Tel: 800 819 3336Tel: (+86) 21 6141 5566Fax: (+86) 21 6141 5567

Czech RepublicSIGMA-ALDRICH spol. s r. o.Tel: (+420) 246 003 200Fax: (+420) 246 003 291

DenmarkSIGMA-ALDRICH DENMARK A/STel: (+45) 43 56 59 10Fax: (+45) 43 56 59 05

FinlandSIGMA-ALDRICH FINLAND OYTel: (+358) 9 350 9250Fax: (+358) 9 350 92555

FranceSIGMA-ALDRICH CHIMIE S.à.r.l.Free Tel: 0800 211 408Free Fax: 0800 031 052Tel: (+33) 474 82 28 00Fax: (+33) 474 95 68 08

GermanySIGMA-ALDRICH CHEMIE GmbHFree Tel: 0800 51 55 000Free Fax: 0800 64 90 000Tel: (+49) 89 6513 0Fax: (+49) 89 6513 1160

GreeceSIGMA-ALDRICH (O.M.) LTD.Tel: (+30) 210 994 8010Fax: (+30) 210 994 3831

HungarySIGMA-ALDRICH KftIngyenes zöld telefon: 06 80 355 355Ingyenes zöld fax: 06 80 344 344Tel: (+36) 1 235 9055Fax: (+36) 1 235 9050

IndiaSIGMA-ALDRICH CHEMICALS PRIVATE LIMITEDTelephoneBangalore: (+91) 80 6621 9600New Delhi: (+91) 11 4165 4255Mumbai: (+91) 22 2570 2364Hyderabad: (+91) 40 4015 5488FaxBangalore: (+91) 80 6621 9650New Delhi: (+91) 11 4165 4266Mumbai: (+91) 22 2579 7589Hyderabad: (+91) 40 4015 5466

IrelandSIGMA-ALDRICH IRELAND LTD.Free Tel: 1800 200 888Free Fax: 1800 600 222Tel: (+353) 1 404 1900Fax: (+353) 1 404 1910

IsraelSIGMA-ALDRICH ISRAEL LTD.Free Tel: 1 800 70 2222Tel: (+972) 8 948 4100Fax: (+972) 8 948 4200

ItalySIGMA-ALDRICH S.r.l.Numero Verde: 800 827018Tel: (+39) 02 3341 7310Fax: (+39) 02 3801 0737

Japan

SIGMA-ALDRICH JAPAN K.K.

Tel: (+81) 3 5796 7300

Fax: (+81) 3 5796 7315

Korea

SIGMA-ALDRICH KOREA

Free Tel: (+82) 80 023 7111

Free Fax: (+82) 80 023 8111

Tel: (+82) 31 329 9000

Fax: (+82) 31 329 9090

Malaysia

SIGMA-ALDRICH (M) SDN. BHD

Tel: (+60) 3 5635 3321

Fax: (+60) 3 5635 4116

Mexico

SIGMA-ALDRICH QUÍMICA, S.A. de C.V.

Free Tel: 01 800 007 5300

Free Fax: 01 800 712 9920

Tel: 52 722 276 1600

Fax: 52 722 276 1601

The Netherlands

SIGMA-ALDRICH CHEMIE BV

Free Tel: 0800 022 9088

Free Fax: 0800 022 9089

Tel: (+31) 78 620 5411

Fax: (+31) 78 620 5421

New Zealand

SIGMA-ALDRICH NEW ZEALAND LTD.

Free Tel: 0800 936 666

Free Fax: 0800 937 777

Tel: (+61) 2 9841 0555

Fax: (+61) 2 9841 0500

Norway

SIGMA-ALDRICH NORWAY AS

Tel: (+47) 23 17 60 60

Fax: (+47) 23 17 60 50

Poland

SIGMA-ALDRICH Sp. z o.o.

Tel: (+48) 61 829 01 00

Fax: (+48) 61 829 01 20

Portugal

SIGMA-ALDRICH QUÍMICA, S.A.

Free Tel: 800 202 180

Free Fax: 800 202 178

Tel: (+351) 21 924 2555

Fax: (+351) 21 924 2610

RussiaSIGMA-ALDRICH RUS, LLCTel: +7 (495) 621 6037 +7 (495) 621 5828Fax: +7 (495) 621 5923

SingaporeSIGMA-ALDRICH PTE. LTD.Tel: (+65) 6779 1200Fax: (+65) 6779 1822

South AfricaSIGMA-ALDRICH SOUTH AFRICA (PTY) LTD.Free Tel: 0800 1100 75Free Fax: 0800 1100 79Tel: (+27) 11 979 1188Fax: (+27) 11 979 1119

SpainSIGMA-ALDRICH QUÍMICA, S.A.Free Tel: 900 101 376Free Fax: 900 102 028Tel: (+34) 91 661 99 77Fax: (+34) 91 661 96 42

SwedenSIGMA-ALDRICH SWEDEN ABTel: (+46) 8 742 4200Fax: (+46) 8 742 4243

SwitzerlandSIGMA-ALDRICH CHEMIE GmbHFree Tel: 0800 80 00 80Free Fax: 0800 80 00 81Tel: (+41) 81 755 2828Fax: (+41) 81 755 2815

United KingdomSIGMA-ALDRICH COMPANY LTD.Free Tel: 0800 717 181Free Fax: 0800 378 785Tel: (+44) 1747 833 000Fax: (+44) 1747 833 313SAFC (UK) Free Tel: 01202 712305

United StatesSIGMA-ALDRICHP.O. Box 14508St. Louis, Missouri 63178Toll-Free: 800 325 3010Toll-Free Fax: 800 325 5052Call Collect: (+1) 314 771 5750Tel: (+1) 314 771 5765Fax: (+1) 314 771 5757

Internetsigma-aldrich.com

Accelerating Customers’ Success

through Leadership in Life Science,

High Technology and Service

Glyco

bio

log

y An

alysis Man

ual

sigma-aldrich.com

Puzzled by Glycobiology?

Our Innovation, Your Research — Shaping the Future of Life Science

2nd edition

Tools for Glycoproteomics and Glycomics

Glycan Labeling and Analysis

Glycoprotein Purification and Detection

Chemical and Enzymatic Deglycosylation

Enzymatic Synthesis and Degradation

Glycobiology Analysis Manual

JFY02060-4013200127





©2007 Sigma-Aldrich Co. All rights reserved. SIGMA, , SAFC, , SIGMA-ALDRICH, ALDRICH, , FLUKA, , and SUPELCO, are trademarks belonging to Sigma-Aldrich Co. and its affiliate Sigma-Aldrich Biotechnology, L.P. Sigma brand products are sold through Sigma-Aldrich, Inc. Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip.

ArgentinaSIGMA-ALDRICH DE ARGENTINA S.A.Free Tel: 0810 888 7446Tel: (+54) 11 4556 1472Fax: (+54) 11 4552 1698

AustraliaSIGMA-ALDRICH PTY LTD.Free Tel: 1800 800 097 Free Fax: 1800 800 096Tel: (+61) 2 9841 0555Fax: (+61) 2 9841 0500

AustriaSIGMA-ALDRICH HANDELS GmbHTel: (+43) 1 605 81 10Fax: (+43) 1 605 81 20

BelgiumSIGMA-ALDRICH NV/SA.Free Tel: 0800 14747Free Fax: 0800 14745Tel: (+32) 3 899 13 01Fax: (+32) 3 899 13 11

BrazilSIGMA-ALDRICH BRASIL LTDA.Free Tel: 0800 701 7425Tel: (+55) 11 3732 3100Fax: (+55) 11 5522 9895

CanadaSIGMA-ALDRICH CANADA LTD.Free Tel: 1800 565 1400Free Fax: 1800 265 3858Tel: (+1) 905 829 9500Fax: (+1) 905 829 9292

ChinaSIGMA-ALDRICH (SHANGHAI) TRADING CO. LTD.Free Tel: 800 819 3336Tel: (+86) 21 6141 5566Fax: (+86) 21 6141 5567

Czech RepublicSIGMA-ALDRICH spol. s r. o.Tel: (+420) 246 003 200Fax: (+420) 246 003 291

DenmarkSIGMA-ALDRICH DENMARK A/STel: (+45) 43 56 59 10Fax: (+45) 43 56 59 05

FinlandSIGMA-ALDRICH FINLAND OYTel: (+358) 9 350 9250Fax: (+358) 9 350 92555

FranceSIGMA-ALDRICH CHIMIE S.à.r.l.Free Tel: 0800 211 408Free Fax: 0800 031 052Tel: (+33) 474 82 28 00Fax: (+33) 474 95 68 08

GermanySIGMA-ALDRICH CHEMIE GmbHFree Tel: 0800 51 55 000Free Fax: 0800 64 90 000Tel: (+49) 89 6513 0Fax: (+49) 89 6513 1160

GreeceSIGMA-ALDRICH (O.M.) LTD.Tel: (+30) 210 994 8010Fax: (+30) 210 994 3831

HungarySIGMA-ALDRICH KftIngyenes zöld telefon: 06 80 355 355Ingyenes zöld fax: 06 80 344 344Tel: (+36) 1 235 9055Fax: (+36) 1 235 9050

IndiaSIGMA-ALDRICH CHEMICALS PRIVATE LIMITEDTelephoneBangalore: (+91) 80 6621 9600New Delhi: (+91) 11 4165 4255Mumbai: (+91) 22 2570 2364Hyderabad: (+91) 40 4015 5488FaxBangalore: (+91) 80 6621 9650New Delhi: (+91) 11 4165 4266Mumbai: (+91) 22 2579 7589Hyderabad: (+91) 40 4015 5466

IrelandSIGMA-ALDRICH IRELAND LTD.Free Tel: 1800 200 888Free Fax: 1800 600 222Tel: (+353) 1 404 1900Fax: (+353) 1 404 1910

IsraelSIGMA-ALDRICH ISRAEL LTD.Free Tel: 1 800 70 2222Tel: (+972) 8 948 4100Fax: (+972) 8 948 4200

ItalySIGMA-ALDRICH S.r.l.Numero Verde: 800 827018Tel: (+39) 02 3341 7310Fax: (+39) 02 3801 0737

Japan

SIGMA-ALDRICH JAPAN K.K.

Tel: (+81) 3 5796 7300

Fax: (+81) 3 5796 7315

Korea

SIGMA-ALDRICH KOREA

Free Tel: (+82) 80 023 7111

Free Fax: (+82) 80 023 8111

Tel: (+82) 31 329 9000

Fax: (+82) 31 329 9090

Malaysia

SIGMA-ALDRICH (M) SDN. BHD

Tel: (+60) 3 5635 3321

Fax: (+60) 3 5635 4116

Mexico

SIGMA-ALDRICH QUÍMICA, S.A. de C.V.

Free Tel: 01 800 007 5300

Free Fax: 01 800 712 9920

Tel: 52 722 276 1600

Fax: 52 722 276 1601

The Netherlands

SIGMA-ALDRICH CHEMIE BV

Free Tel: 0800 022 9088

Free Fax: 0800 022 9089

Tel: (+31) 78 620 5411

Fax: (+31) 78 620 5421

New Zealand

SIGMA-ALDRICH NEW ZEALAND LTD.

Free Tel: 0800 936 666

Free Fax: 0800 937 777

Tel: (+61) 2 9841 0555

Fax: (+61) 2 9841 0500

Norway

SIGMA-ALDRICH NORWAY AS

Tel: (+47) 23 17 60 60

Fax: (+47) 23 17 60 50

Poland

SIGMA-ALDRICH Sp. z o.o.

Tel: (+48) 61 829 01 00

Fax: (+48) 61 829 01 20

Portugal

SIGMA-ALDRICH QUÍMICA, S.A.

Free Tel: 800 202 180

Free Fax: 800 202 178

Tel: (+351) 21 924 2555

Fax: (+351) 21 924 2610

RussiaSIGMA-ALDRICH RUS, LLCTel: +7 (495) 621 6037 +7 (495) 621 5828Fax: +7 (495) 621 5923

SingaporeSIGMA-ALDRICH PTE. LTD.Tel: (+65) 6779 1200Fax: (+65) 6779 1822

South AfricaSIGMA-ALDRICH SOUTH AFRICA (PTY) LTD.Free Tel: 0800 1100 75Free Fax: 0800 1100 79Tel: (+27) 11 979 1188Fax: (+27) 11 979 1119

SpainSIGMA-ALDRICH QUÍMICA, S.A.Free Tel: 900 101 376Free Fax: 900 102 028Tel: (+34) 91 661 99 77Fax: (+34) 91 661 96 42

SwedenSIGMA-ALDRICH SWEDEN ABTel: (+46) 8 742 4200Fax: (+46) 8 742 4243

SwitzerlandSIGMA-ALDRICH CHEMIE GmbHFree Tel: 0800 80 00 80Free Fax: 0800 80 00 81Tel: (+41) 81 755 2828Fax: (+41) 81 755 2815

United KingdomSIGMA-ALDRICH COMPANY LTD.Free Tel: 0800 717 181Free Fax: 0800 378 785Tel: (+44) 1747 833 000Fax: (+44) 1747 833 313SAFC (UK) Free Tel: 01202 712305

United StatesSIGMA-ALDRICHP.O. Box 14508St. Louis, Missouri 63178Toll-Free: 800 325 3010Toll-Free Fax: 800 325 5052Call Collect: (+1) 314 771 5750Tel: (+1) 314 771 5765Fax: (+1) 314 771 5757

Internetsigma-aldrich.com

Accelerating Customers’ Success

through Leadership in Life Science,

High Technology and Service

Glyco

bio

log

y An

alysis Man

ual

Visit sigma.com/glycomanual and request your copy.

Whether you are an expert in carbohydrate biology and chemistry or just getting started in glycomics, the Glycobiology Analysis Manual provides the products and methods you need to solve your glycomics puzzle!

• Innovative products and kits, including GlycoProfile™ metabolic labeling reagents for glycomics expression, glycosyltransferases for glycan synthesis, and kits for chemical and enzymatic deglycosylation

• Updated and expanded technical content on glycan detection labeling, mass spectrometry, NMR spectroscopy, and other key techniques

• Structural and functional reviews of glycoproteins, proteoglycans, glycosphingolipids, and other carbohydrate-modified biomolecules

GlycoProfile is a trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co.



















http://www.sigma-aldrich.com

http://www.sigma.com/glycomanual


Dex

tran

an

d R

elat

ed P

oly

sacc

har

ides

Name Mol Wt Cat. No.


BioChemika

Dextran from Leuconostoc spp. Mr ~2,000,000 95771-10G95771-50G95771-250G

BioChemika

Dextran from Leuconostoc mesenteroides average mol wt 9,000-11,000 D9260-10GD9260-50GD9260-100GD9260-500G

Dextran from Leuconostoc mesenteroides average mol wt 35,000-45,000 D1662-10GD1662-50GD1662-100GD1662-500G

Dextran from Leuconostoc mesenteroides average mol wt 60,000-90,000 D3759-1KGD3759-2KG

Dextran from Leuconostoc mesenteroides Mr ~60,000 31397-100G31397-500G

BioChemika

Dextran from Leuconostoc mesenteroides average mol wt 64,000-76,000 D4751-10GD4751-50GD4751-100GD4751-500GD4751-1KG

Dextran from Leuconostoc mesenteroides average mol wt 100,000-200,000 D4876-50GD4876-100GD4876-500GD4876-1KG

Dextran from Leuconostoc mesenteroides Mr ~200,000 31398-25G31398-100G31398-500G

BioChemika

Dextran from Leuconostoc mesenteroides average mol wt 425,000-575,000 D1037-50GD1037-100GD1037-500G

Dextran from Leuconostoc mesenteroides average mol wt ~2,000,000 D5376-100GD5376-500G

Dextran from Leuconostoc mesenteroides average mol wt 5,000,000-40,000,000 D5501-100GD5501-500GD5501-1KG

industrial grade

DextranFor General Applications, continued









http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=D9260&Brand=SIAL

































Dextran

and

Related

Polysacch

arides

For Use as Standards in Gel Permeation Chromatography (GPC)Certified dextran standards have been certified by the Deutsches Institut für Normung e.V. (DIN, Berlin, Germany).

Name Description Mn MP MW Cat. No.

Dextran standard 1,000 certified according to DIN ~1,010 ~1,080 ~1,270 00268-500MG

BioChemika

Dextran standard 5,000 certified according to DIN ~3,260 ~4,440 ~5,220 00269-100MG00269-500MG

BioChemika


BioChemika


BioChemika


BioChemika


BioChemika


BioChemika

Dextran standard 270,000 certified according to DIN ~164200 ~196,300 ~273,000 00894-100MG00894-500MG

BioChemika


BioChemika


BioChemika

Dextran from Leuconostoc mesenteroides standard 1,000 ~1,000 ~1,100 ~1,300 31416-100MG

BioChemika

Dextran from Leuconostoc mesenteroides standard 5,000 ~3,300 ~4,400 ~5,200 31417-100MG-F

BioChemika


BioChemika


BioChemika


BioChemika


BioChemika


BioChemika


BioChemika


BioChemika


BioChemika

Dextran from Leuconostoc mesenteroides standard 1,400,000 ~65,100 ~1,223,000 ~1,394,000 49297-100MG-F

BioChemika




























Dex

tran

an

d R

elat

ed P

oly

sacc

har

ides

Dextran Sulfate

Name Mol. Wt. Additives Cat. No.

Dextran sulfate sodium salt from Leuconostoc spp. Mr 5,000 - 31404-5G-F31404-25G-F

BioChemika

Dextran sulfate sodium salt from Leuconostoc spp. 6,500-10,000 - D4911-1GD4911-10GD4911-50GD4911-100G

Dextran sulfate sodium salt from Leuconostoc spp. average mol wt 9,000-20,000 - D6924-1GD6924-10GD6924-50G

Dextran sulfate sodium salt from Leuconostoc spp. Mr 100,000 - 66786-50G-F

BioChemika

Dextran sulfate sodium salt from Leuconostoc spp. average mol wt >500,000 (dextran starting material)

phosphate buffer, pH 6-8 0.5-2.0%

D6001-1GD6001-10GD6001-50GD6001-100GD6001-500G

Dextran sulfate sodium salt from Leuconostoc spp. average Mw >500,000 (dextran starting material)

phosphate buffer 0.5-2%

D8906-5GD8906-10GD8906-50GD8906-100GD8906-500G

for molecular biology

Immobilized Dextran

Name Synonyms Particle Size (μm) Cat. No.

CM-Dextran sodium salt CM-D; Carboxymethyl-dextran - 86524-10G-F86524-50G-F86524-100G-F86524-250G-F86524-500G-F

BioChemika

DEAE-Dextran hydrochloride Diethylaminoethyl-dextran hydrochloride - D9885-10GD9885-50GD9885-100G

DEAE-Dextran hydrochloride Diethylaminoethyl-dextran hydrochloride - 30461-25G

BioChemikafor molecular biology

Dextran cross-linked G-25 - 50 - 150 01468-250G-F

BioChemika

Dextran cross-linked G-50 - 20 - 80 94504-50G-F94504-250G-F

BioChemika

Dextran cross-linked G-50 - 50 - 150 40359-50G-F40359-250G-F

BioChemika

Dextran cross-linked G-50 - 100 - 300 68263-10G-F68263-50G-F68263-250G-F

BioChemika









































Dextran

and

Related

Polysacch

arides

Dextrins and PullulansName Type Product Line, Grade Impurities Cat. No.

Dextrin from corn Type I - Reducing sugar ≤5% D2006-100GD2006-500GD2006-1KG

This grade is prepared by removing a minimum of 95% of the water-insolubles, and almost all of the alcohol-solubles.

Dextrin from corn Type II commercial grade - D2131-500GD2131-2.5KG

Dextrin from corn Type III commercial grade - D2256-500GD2256-2.5KG

Dextrin from maize starch - BioChemika reducing matter in dry substance ~10% 31410-100G31410-500G

Dextrin from maize starch - BioChemika reducing matter in dry substance ~20% 31414-100G31414-500G

Dextrin from potato starch Type IV - Reducing sugar ≤5% D4894-500GD4894-1KG

Dextrin from potato starch - BioChemika, for microbiology - 31400-250G31400-1KG

Pullulan from Aureobasidium pullulans - - - P4516-1GP4516-5GP4516-25G

substrate for pullulanase suitable

Pullulan Standard 340[9057-02-7]

for GPCMn ~342

Mp ~342

Mw ~342

Mw/Mn ~1.00

exact values can be taken from the accompanying certificate of analysis

76651-100MG 100 mg

Pullulan Standard Set[9057-02-7]

for GPC, Mp 342-710’000exact values can be taken from the accompanying certificate of analysis

Set contains 10 different Pullulan Standards with 0.1 g with Mw ~320, ~1’300, ~6’000, ~12’000, ~22’000, ~50’000, ~110’000, ~200’000, ~400’000, ~800’000

96351-1KT 1 kit

Dextran[9004-54-0] [C6H10O5]n

BioChemika, for GPC, standard-Set Mp 1,000-400,000exact values can be taken from the accompanying certificate of analysis

set contains 10 different dextran standards with 500 mg with Mp ~1,100 (Fluka 31416), ~4,400 (Fluka 31417), ~10,000 (Fluka 31418), ~20,000 (Fluka 31419), ~45,000 (Fluka 31420),~65,000 (Fluka 31421), ~125,000 (Fluka 31422), ~195,000 (Fluka 31423), ~275,000 (Fluka 31424), ~400,000 (Fluka 31425); The components can also be ordered separately under the Fluka product numbers indicated.

31430-1EA 1 set

Pullulan Standard 180[9057-02-7]

for GPCMn ~180

Mp ~180

Mw ~180

Mw/Mn ~1.00

exact values can be taken from the accompanying certificate of analysis

90950-100MG 100 mg

Additional Gel Permeation Chromatography (GPC) Standards and Sets
























Gly

coPr

ofi

le™

β-E

limin

atio

n K

it

O-Linked glycans are linked through the hydroxyl of Ser (serine) or Thr (threonine) and do not require a consensus sequence, although a number of core structures have been identified. Despite their complexity and apparent lack of uniformity, O-linked glycans are being studied more frequently in recent research. This is due to the increased awareness of their multiple roles and importance in biological function and disease. In order to further understand O-linked glycans, the glycan is often removed from the protein for analysis.

Enzymatic methods of O-linked deglycosylation include sequentially hydrolyzing sugars by use of a series of exoglycosidases, followed by O-glycosidase. Although effective at removing the glycan, this process is tedious and does not result in an intact glycan. There are a number of chemical methods used to remove O-glycans. These include hydrazinolysis and trifluoromethanesulfonic (TFMS) acid, which remove both N- and O-linked glycans, and can be destructive to the glycans, thus preventing accurate analysis.

Alkaline β-elimination is a chemical method of deglycosylation commonly used to specifically remove O-glycans. Traditionally, this method uses a combination of sodium hydroxide and sodium borohydride. The O-glycan linkage is easily hydrolyzed using dilute alkaline solution under mild conditions. The presence of a reducing agent can keep the glycan from “peeling” after being released, but the process significantly degrades the protein or peptide. Other methods, such as using sodium hydroxide alone or with borane-ammonia, also easily hydrolyze the linkage, but are not very efficient at keeping both moieties intact.

A novel non-reductive β-elimination kit has been developed that keeps both protein and glycan intact. The method is much easier to use than the traditional β-elimination methodologies. There is no tedious neutralization of the borohydride or ion exchange chromatography to be performed. This technology allows for complete glycoproteomic analysis of O-linked glycoproteins, as never before possible.

GlycoProfile™ β-Elimination Kit

O-Glycan Removal: A Novel β-Elimination Method

Sigma’s GlycoProfile β-Elimination Kit is unique in that it does not completely destroy the protein, as seen with other traditional methods of β-elimination. SDS-PAGE of proteins before and after deglycosylation. Gel has been stained with EZBlue™ Gel Staining Reagent (Cat. No. G1041) and destained with water.

1 2 3 4 5 6 7 8 9 10 11 12

Lane 1: ColorBurst™ Marker, High Range (Cat. No. C1992) Lanes 2 and 3: Fetuin prior to β-elimination Lanes 4 and 5: Fetuin incubated in 50 mM NaOH overnight Lanes 6 and 7: Fetuin incubated in NaBH4 and NaOH overnight (traditional β−elimination) Lanes 8 and 9: Fetuin incubated in Sigma’s β−elimination reagent at 4-8° C overnight Lanes 10 and 11: Fetuin incubated in Sigma’s β−elimination reagent at room temperature overnight Lane 12: SigmaMarker™, Wide Range (Cat. No. S8445)





http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=S8445&Brand=SIGMA


sigma-aldrich.com

Glyco

Profile

™ β

-Elimin

ation

Kit

Sample# of Unique

Peptides% Sequence

Coverage

TrypZean, before deglycosylation* 24 55

TrypZean, after deglycosylation 29 55

Glycophorin A, before deglycosylation 17 53

Glycophorin A, after deglycosylation 22 59

* Trypzean is recombinant trypsin, bovine sequence, expressed in corn.

GlycoProfile™ β-Elimination Kit 8

Components

β-Elimination Reagent 940 μLSodium hydroxide (Sigma S8263) 60 μLProtein Enrichment Column 10 eachGlycan Enrichment Column 10 each store at: Room temp

PP0540-1KT 1 kit

Navigate!

Let Sigma help navigate your proteomics research.

■ Full range of progressive technologies for biomarker discovery■ Proteomic standards, calibrants, and markers■ Intuitive design tools for peptide libraries

Order your Proteomics Product Guide today. Visit sigma.com/pepguide

TrypZean, BEFORE DEGLYCOSYLATION

TRY1_BOVIN (100%), 25,424.8 DaCationic trypsin precursor (EC 3.4.21.4) (Beta-Trypsin) [Contains: Alpha-trypsin chain 2] (Fragment) – Bos taurus (Bovine)24 unique peptides, 25 unique spectra, 463 total spectra, 134/243 amino acids (55% sequence coverage)

FIFLALLGAAVAFPVDDDDKIVGGYTCGANTVPYQVSLNSGYHFCGGSLINSQWVVSAAHCYKSGIQVRLGEDNINVVEGNEQFISASKSIVHPSYNSNTLNNDIMLIKLKSAASLNSRVASISLPTSCASAGTQCLISGWGNTKSSGTSYPDVLKCLKAPILSDSSCKSAYPGQITSNMFCAGYLEGGKDSCQGDSGGPVVCSGKLQGIVSWGSGCAQKNKPGVYTKVCNYVSWIKQTIASN

TrypZean, AFTER DEGLYCOSYLATION

TRY1_BOVIN (100%), 25,424.8 DaCationic trypsin precursor (EC 3.4.21.4) (Beta-Trypsin) [Contains: Alpha-trypsin chain 2] (Fragment) – Bos taurus (Bovine)29 unique peptides, 30 unique spectra, 573 total spectra, 134/243 amino acids (55% sequence coverage)

FIFLALLGAAVAFPVDDDDKIVGGYTCGANTVPYQVSLNSGYHFCGGSLINSQWVVSAAHCYKSGIQVRLGEDNINVVEGNEQFISASKSIVHPSYNSNTLNNDIMLIKLKSAASLNSRVASISLPTSCASAGTQCLISGWGNTKSSGTSYPDVLKCLKAPILSDSSCKSAYPGQITSNMFCAGYLEGGKDSCQGDSGGPVVCSGKLQGIVSWGSGCAQKNKPGVYTKVCNYVSWIKQTIASN

GLYCOPHORIN A, BEFORE DEGLYCOSYLATION

GLPA_HUMAN (100%), 16,273.7 DaGlycophorin A precursor (PAS-2) (Sialoglycoprotein alpha) (MN sialoglycoprotein) (CD235a antigen) – Homo sapiens (Human)17 unique peptides, 18 unique spectra, 79/150 amino acids (53% coverage)

MYGKIIFVLLLSAIVSISASSTTGVAMHTSTSSSVTKSYISSQTNDTHKRDTYAATPRAHEVSEISVRTVYPPEEETGERVQLAHHFSEPEITLIIFGVMAGVIGTILLISYGIRRLIKKSPSDVKPLPSPDTDVPLSSVEIENPETSDQ

GLYCOPHORIN A, AFTER DEGLYCOSYLATION

GLPA_HUMAN (100%), 16,273.7 DaGlycophorin A precursor (PAS-2) (Sialoglycoprotein alpha) (MN sialoglycoprotein) (CD235a antigen) – Homo sapiens (Human)22 unique peptides, 25 unique spectra, 89/150 amino acids (59% coverage)

MYGKIIFVLLLSAIVSISASSTTGVAMHTSTSSSVTKSYISSQTNDTHKRDTYAATPRAHEVSEISVRTVYPPEEETGERVQLAHHFSEPEITLIIFGVMAGVIGTILLISYGIRRLIKKSPSDVKPLPSPDTDVPLSSVEIENPETSDQ

The GlycoProfile β-Elimination Kit allows for proteomic analysis of O-linked glycoproteins after removal of the O-glycan. The following data is summarized from LC-MS analysis of the tryptic peptides of two glycoproteins (TrypZean™ recombinant trypsin, and glycophorin A) before and after deglycosylation.

http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=PP0540&Brand=SIGMA

http://www.sigma.com/pepguide


Gly

coPr

ofi

le™

β-E

limin

atio

n K

it

Related Kits for Deglycosylation

GlycoProfile™ IV Chemical Deglycosylation Kit

TFMS Deglycosylation SystemThe GlycoProfile™ IV Kit utilizes trifluoromethanesulfonic acid (TFMS) in a deglycosylation system that completely removes all N- and O-linked glycans while preserving the protein/polypeptide structure. TFMS hydrolysis of glycoproteins results in minimal protein degradation while the released glycans are destroyed. For high molecular weight or complex, non-mammalian glycoproteins, an optional scavenger species is included. Complete deglycosylation of glycoproteins takes place in as little as 30 minutes. store at: 2-8°C

PP0510-1KT 1 kit

GlycoProfile™ I, In-Gel Deglycosylation Kit

The GlycoProfile™ Enzymatic In-gel N-Deglycosylation Kit is optimized to provide a convenient, reproducible method to N-deglycosylate and digest protein samples from one dimensional (1D) and two dimensional (2D) polyacrylamide gel pieces for subsequent mass spectrometry or HPLC analysis.

This procedure is suitable for Coomassie® Brilliant Blue and colloidal Coomassie stained gels. Silver stained gels may also be used if properly destained. store at: 2-8°C

PP0200-1KT 1 kit

Glycoprofile™ II, Enzymatic In-Solution N-Deglycosylation Kit

GlycoProfile™ II has been optimized to provide a convenient and reproducible method to remove

N-linked glycans from glycoproteins and is compatible with subsequent MALDI-TOF mass spectrometric analysis without interference from any of the reaction components.

Contains sufficient reagents for a minimum of 20 reactions when the sample size is between one to two mg of a typical glycoprotein. store at: 2-8°C

PP0201-1KT 1 kit

Enzymatic Protein Deglycosylation Kit

Contains all enzymes and reagents needed to completely remove all N- and simple O-linked including polysialylated carbohydrates from glycoproteins as well as additional enzymes & reagents needed to remove complex O-linked carbohydrates.ship: wet ice store at: 2-8°C

EDEGLY-1KT 1 kit

Native Protein Deglycosylation Kit

The Native Protein Deglycosylation is designed for the deglycoslylation of N-linked oligosaccharides from PNGase F-resistant native proteins. Endoglycosidases F1, F2, and F3 are less sensitive to protein conformation than PNGase F and are more suitable for removal of all classes of N-linked oligosaccharides without protein denaturation.ship: wet ice store at: 2-8°C

NDEGLY-1KT 1 kit

sigma-aldrich.com

Your Favorite Gene is a search tool that matches your gene of interest against thousands of research products available from Sigma.

■ Flexible search options:

— Use a wide variety of terms to locate detailed gene information

— Refine or expand your search by pathways, function & more

■ Unparalleled product coverage matched to your gene of interest:

— 150,000 shRNAs

— 2,000 Bioactive small molecules

— 6,000 Antibodies, proteins & kits

— 725,000 siRNAs (including 700 validated)

Search Your Favorite Gene at sigma.com/yfg

Your comprehensive gene search tool from Sigma

TM






http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=EDEGLY&Brand=SIGMA

http://www.sigmaaldrich.com/catalog/search/ProductDetail?ProdNo=NDEGLY&Brand=SIGMA

http://www.sigma-aldrich.com

http://www.sigma.com/yfg

Our Innovation, Your Research — Shaping the Future of Life Science

With Highly-validated shRNA Libraries . . .

For more information on MISSION shRNA libraries, visit sigma.com/trc2bio

We’ve Got You Covered.

Your RNAi projects are unique and demand the highest quality reagents for the cell lines most relevant to your research. When you partner with Sigma’s MISSION® shRNA team, you gain access to an extensively validated genome-wide shRNA collection, ideal screening formats, and flexible custom services designed to accelerate your discovery. Save time and money when you take advantage of Sigma’s world-class expertise in high-throughput lentiviral manufacturing and the validation efforts of TRC-2, sponsored in part by Sigma. So, start your shRNA research today. We've got you covered.

MISSION is a registered trademark of Sigma-Aldrich Co. and its affiliate Sigma-Aldrich Biotechnology, L.P.

sigma-aldrich.com

LBK70933-506322

0118





©2008 Sigma-Aldrich Co. All rights reserved. SIGMA, , SAFC, , SIGMA-ALDRICH, ALDRICH, , FLUKA, , and SUPELCO, are trademarks belonging to Sigma-Aldrich Co. and its affiliate Sigma-Aldrich Biotechnology, L.P. Sigma brand products are sold through Sigma-Aldrich, Inc. Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip. Eppendorf is a registered trademark of Eppendorf-Netheler-Hinz GmbH. Sephadex and Sepharose are registered trademarks of GE Healthcare. Coomassie is a registered trademark of Imperial Chemical Industries Ltd. ColorBurst, EZBlue, GlycoProfile, Prestige Antibodies, ProteoSilver, and SigmaMarker are trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co. MISSION and TraceSELECT are registered trademarks of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co. Triton is a registered trademark of Union Carbide Corp. TrypZean is a trademark of ProdiGene, Inc. ARIXTRA is a registered trademark of the GlaxoSmithKline group of companies.

Sigma-Aldrich3050 Spruce StreetSt. Louis, MO 63103 USAsigma-aldrich.com

Accelerating Customers’

Success through Leadership

in Life Science, High

Technology and Service

ArgentinaSIGMA-ALDRICH DE ARGENTINA S.A. Free Tel: 0810 888 7446 Tel: (+54) 11 4556 1472 Fax: (+54) 11 4552 1698

AustraliaSIGMA-ALDRICH PTY LTD. Free Tel: 1800 800 097 Free Fax: 1800 800 096 Tel: (+61) 2 9841 0555 Fax: (+61) 2 9841 0500

AustriaSIGMA-ALDRICH HANDELS GmbH Tel: (+43) 1 605 81 10 Fax: (+43) 1 605 81 20

BelgiumSIGMA-ALDRICH NV/S.A.Free Tel: 0800 14747 Free Fax: 0800 14745 Tel: (+32) 3 899 13 01 Fax: (+32) 3 899 13 11

BrazilSIGMA-ALDRICH BRASIL LTDA.Free Tel: 0800 701 7425 Tel: (+55) 11 3732 3100 Fax: (+55) 11 5522 9895

CanadaSIGMA-ALDRICH CANADA LTD.Free Tel: 1800 565 1400 Free Fax: 1800 265 3858 Tel: (+1) 905 829 9500 Fax: (+1) 905 829 9292

ChinaSIGMA-ALDRICH (SHANGHAI) TRADING CO. LTD.Free Tel: 800 819 3336 Tel: (+86) 21 6141 5566 Fax: (+86) 21 6141 5567

Czech RepublicSIGMA-ALDRICH spol. s r. o. Tel: (+420) 246 003 200 Fax: (+420) 246 003 291

DenmarkSIGMA-ALDRICH DENMARK A/S Tel: (+45) 43 56 59 10 Fax: (+45) 43 56 59 05

FinlandSIGMA-ALDRICH FINLAND OYTel: (+358) 9 350 9250 Fax: (+358) 9 350 92555

FranceSIGMA-ALDRICH CHIMIE S.à.r.l. Free Tel: 0800 211 408Free Fax: 0800 031 052Tel: (+33) 474 82 28 00 Fax: (+33) 474 95 68 08

GermanySIGMA-ALDRICH CHEMIE GmbHFree Tel: 0800 51 55 000 Free Fax: 0800 64 90 000 Tel: (+49) 89 6513 0 Fax: (+49) 89 6513 1160

GreeceSIGMA-ALDRICH (O.M.) LTD.Tel: (+30) 210 994 8010 Fax: (+30) 210 994 3831

HungarySIGMA-ALDRICH Kft Ingyenes telefonszám: 06 80 355 355 Ingyenes fax szám: 06 80 344 344 Tel: (+36) 1 235 9055 Fax: (+36) 1 235 9050

IndiaSIGMA-ALDRICH CHEMICALS PRIVATE LIMITED Telephone Bangalore: (+91) 80 6621 9600 New Delhi: (+91) 11 4358 8000 Mumbai: (+91) 22 2570 2364 Hyderabad: (+91) 40 4015 5488 Fax Bangalore: (+91) 80 6621 9650 New Delhi: (+91) 11 4358 8001 Mumbai: (+91) 22 2579 7589 Hyderabad: (+91) 40 4015 5466

IrelandSIGMA-ALDRICH IRELAND LTD. Free Tel: 1800 200 888 Free Fax: 1800 600 222Tel: +353 (0) 402 20370Fax: + 353 (0) 402 20375

IsraelSIGMA-ALDRICH ISRAEL LTD.Free Tel: 1 800 70 2222 Tel: (+972) 8 948 4100 Fax: (+972) 8 948 4200

ItalySIGMA-ALDRICH S.r.l. Numero Verde: 800 827018 Tel: (+39) 02 3341 7310 Fax: (+39) 02 3801 0737

JapanSIGMA-ALDRICH JAPAN K.K. Tel: (+81) 3 5796 7300 Fax: (+81) 3 5796 7315

KoreaSIGMA-ALDRICH KOREA Free Tel: (+82) 80 023 7111 Free Fax: (+82) 80 023 8111 Tel: (+82) 31 329 9000 Fax: (+82) 31 329 9090

MalaysiaSIGMA-ALDRICH (M) SDN. BHDTel: (+60) 3 5635 3321 Fax: (+60) 3 5635 4116

MexicoSIGMA-ALDRICH QUÍMICA, S.A. de C.V.Free Tel: 01 800 007 5300 Free Fax: 01 800 712 9920 Tel: 52 722 276 1600 Fax: 52 722 276 1601

The NetherlandsSIGMA-ALDRICH CHEMIE BVFree Tel: 0800 022 9088 Free Fax: 0800 022 9089 Tel: (+31) 78 620 5411 Fax: (+31) 78 620 5421

New ZealandSIGMA-ALDRICH NEW ZEALAND LTD. Free Tel: 0800 936 666 Free Fax: 0800 937 777 Tel: (+61) 2 9841 0555 Fax: (+61) 2 9841 0500

NorwaySIGMA-ALDRICH NORWAY AS Tel: (+47) 23 17 60 60 Fax: (+47) 23 17 60 50

PolandSIGMA-ALDRICH Sp. z o.o. Tel: (+48) 61 829 01 00 Fax: (+48) 61 829 01 20

PortugalSIGMA-ALDRICH QUÍMICA, S.A.Free Tel: 800 202 180 Free Fax: 800 202 178 Tel: (+351) 21 924 2555 Fax: (+351) 21 924 2610

RussiaSIGMA-ALDRICH RUS, LLC Tel: +7 (495) 621 6037 +7 (495) 621 5828 Fax: +7 (495) 621 5923

SingaporeSIGMA-ALDRICH PTE. LTD.Tel: (+65) 6779 1200 Fax: (+65) 6779 1822

SlovakiaSIGMA-ALDRICH spol. s r. o. Tel: (+421) 255 571 562Fax: (+421) 255 571 564

South AfricaSIGMA-ALDRICH SOUTH AFRICA (PTY) LTD.Free Tel: 0800 1100 75 Free Fax: 0800 1100 79 Tel: (+27) 11 979 1188 Fax: (+27) 11 979 1119

SpainSIGMA-ALDRICH QUÍMICA, S.A. Free Tel: 900 101 376 Free Fax: 900 102 028 Tel: (+34) 91 661 99 77 Fax: (+34) 91 661 96 42

SwedenSIGMA-ALDRICH SWEDEN ABTel: (+46) 8 742 4200 Fax: (+46) 8 742 4243

SwitzerlandSIGMA-ALDRICH CHEMIE GmbH Free Tel: 0800 80 00 80 Free Fax: 0800 80 00 81 Tel: (+41) 81 755 2828 Fax: (+41) 81 755 2815

United KingdomSIGMA-ALDRICH COMPANY LTD.Free Tel: 0800 717 181 Free Fax: 0800 378 785 Tel: (+44) 1747 833 000 Fax: (+44) 1747 833 313 SAFC (UK) Tel: 01202 712305

United StatesSIGMA-ALDRICH P.O. Box 14508 St. Louis, Missouri 63178 Toll-Free: 800 325 3010 Toll-Free Fax: 800 325 5052 Call Collect: (+1) 314 771 5750 Tel: (+1) 314 771 5765 Fax: (+1) 314 771 5757

Internet sigma-aldrich.com

biofiles 3

Documents

dextran sulfate

carbohydrate analysis

additional antibodies

carbohydrate analysisstart

heparan sulfate

prestige antibodies

ptm analysis

guidance proteins