Next Generation

Lectins

Recombinant Lectins Ensure Stable Supplies

Ready-to-Use Lectin Solution

For Glycobiology & Cell Biology

Wako has been developing lectins for sugar chain research, aiming to ensure those stable supplies. The first launch is recombinant BC2LCN (AiLecS1) for research of human induced pluripotent stem (iPS) and embryonic stem (ES) cells. rBC2LCN (AiLecS1) is expected to be applicable to not only stem cell research but also to regenerative medicine since it is reportedly a useful marker for detection of undifferentiated cells.

rBC2LCN-FITC DAPI MERGE

Apr. 2016

Application - Detections of an undifferentiation marker of human iPS cells

Wako Pure Chemical Ind., Ltd. www.wako-chem.co.jp

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Fax: 81-6-6203-1999

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■ rBC2LCN (AiLecS1) from cDNA of Burkholderia cenocepacia

OCT3/4

TRA-1-81

rBC2LCN -Cy3

rBC2LCN -FITC

BSA-Cy3

BSA-FITC

DAPI MERGE

rBC2LCN 029-18061 (1mg), 025-18063 (1mg×5)

Glycan specificity Fucα1-2Galβ1-3GalNAc (H type 3) Fucα1-2Galβ1-3GlcNAc (H type 1)

Formulation PBS (pH 7.4)

cDNA source Synthetic cDNA based on Burkholderia cenocepacia

Protein concentration

1.2～1.8mg/ml (BCA Assay)

Source E. coli expressed Burkholderia cenocepacia lectin (BC2LCN/AiLecS1)

Molecular weight (theoretical value)

15,807

Purification Affinity purification by fucose agarose. Storage condition Keep at -20℃ before use. After thawing, keep at 2-10℃. Avoid the freeze-thaw cycle.

Product Outline

Detection of undifferentiated iPS cells using rBC2LCN

rBC2LCN References

rBC2LCN, a marker of undifferentiated human ES・iPS cells

Point in Check

rBC2LCN reportedly recognized an intramolecular sugar chain of Podocalyxin, exclusively, which exposes on the cell surface of undifferentiated human iPS cells. Please see the reference 1).

rBC2LCN Cy3

rBC2LCN FITC

DAPI MERGE BF

SSEA-4

TRA-1-81

＜Detection condition＞ Fluorescent labeled rBC2LCN Final concentration : 0.1-0.5 μg/mL (1w/v% BSA, 1×PBS) Incubation time : 60 min. (RT, in the dark) Cell : Human iPS 201B7 strain Fixing condition: 4w/v% Paraformaldehyde for 10min. at RT or 4℃.

Detection of undifferentiated marker in human iPS cell colony naturally differentiated

Point in Check rBC2LCN （rLectin）

Anti TRA-1-81 Mab （Mouse IgM）

Anti SSEA-4 Mab (Mouse IgG3)

M. W. （kDa） 16 900 150

Flow Cytometry ○ ○ ○

Cytological staining ○ ○ ○

Live cell imaging ○ △ - Non-specific binding (Cytological staining)

Low High Low

1) Tateno, H., Matsushima, A., Hiemori, K., Onuma, Y., Ito, Y., Hasehira, K., Nishimura, K., Ohtaka, M., Takayasu, S., Nakanishi, M., Ikehara, Y., Nakanishi, M., Ohnuma, K., Chan, T., Toyoda, M., Akatsu, H., Umezawa, A., Asashima, M. and Hirabayashi, J. “Podocalyxin is a Glycoprotein ligand of the human pluripotent stem cell-specific probe rBC2LCN.”, Stem Cells Translational Medicine, 2(4), 265-73(2013). 2) Onuma, Y., Tateno, H., Hirabayashi, J., Ito, Y. and Asashima, M. “rBC2LCN, a new probe for live cell imaging of human pluripotent stem cells.”, Biochem. Biophys. Res. Commun., 431(3), 524-9(2013). 3) Tateno, H., Toyota, M., Saito, S., Onuma, Y., Ito, Y., Hiemori, K., Fukumura, M., Matsushima, A., Nakanishi, M., Ohnuma, K., Akutsu, H., Umezawa, A., Horimoto, K., Hirabayashi, J. and Asashima, M. “Glycome diagnosis of human induced pluripotent stem cells using lectin microarray.”, J. Biol. Chem., 286(23), 20345-53(2011). 4) Tateno, H., Onuma, Y., Ito, Y., Hiemori, K., Aiki, Y., Shimizu, M., Higuchi, K., Fukuda, M., Warashina, M., Honda, S., Asashima, M. and Hirabayashi, J. “A medium hyperglycosylated podocalyxin enables noninvasiva and quantitative detection of tumorigenic human pluripotent stem cells.”, Sci. Rep., 4, 4069(2014).

＊1： Lectin group means one kind of classification of expediency based on the glycan specificity. It doesn’t warrant the recognition ability of all glycans.

＊2： The information of the glycan specificity are the common information. These don’t limit the specificity of lectins.

Lectin Groups＊1 Name Code No. Package

Size

cDNA Source

Glycan Specificity ＊2

Fucose-

Complex

BC2LCN

（AiLecS1)

029-18061

025-18063 1mg

1mg×5

cDNA Source : Burkholderia cenocepacia

Target Glycan : Fucα1-2Galβ1-3GalNAc (H type 3), Fucα1-2Galβ1-3GlcNAc (H type 1)

AAL 018-25201

014-25203 1mg

1mg×5

cDNA Source : Aleuria aurantia

Target Glycan : Fucose moieties

PA-IIL 160-26711

166-26713 1mg

1mg×5

cDNA Source : Pseudomonas aeruginosa

Target Glycan : Fucose, Fucose containing oligosaccharides, Mannose

RS-Fuc 183-03101

189-03103 1mg

1mg×5

cDNA Source : Ralstonia solanacearum

Target Glycan : Fucose

Glc/GlcNAc-

Complex

F17AG 062-06281

068-06283 1mg

1mg×5

cDNA Source : E. coli

Target Glycan : GlcNAc

Malectin 136-18221

132-18223 1mg

1mg×5

cDNA Source : Human

Target Glycan : Glc2-N-biose

Gal/GalNAc-

Complex

ABA 015-24851

011-24853 1mg

1mg×5

cDNA Source : Agaricus bisporus

Target Glycan : Galβ1-3GalNAc (T antigen), Galβ1-3GlcNAc (Lacto-N-biose)

CGL2 033-23771

039-23773 1mg

1mg×5

cDNA Source : Coprinopsis cinerea

Target Glycan : βGal, GalNAcα1-3Gal (Blood Group A), Galα1-3Gal (Blood Group B)

CNL 039-23631

035-23633 1mg

1mg×5

cDNA Source : Clitocybe nebularis

Target Glycan : α/βGalNAc, GalNAcβ1-4GlcNAc,

GalNAcα1-3[Fucα1-2]Galβ1-4GlcNAc (Blood Group A)

Discoidin I 045-33541

041-33543 0.5mg

0.5mg×5

cDNA Source : Dictyostelium discoideum

Target Glycan : αGalNAc (Tn antigen)

Discoidin II 042-33431 0.5mg cDNA Source : Dictyostelium discoideum

Target Glycan : Gal, LacNAc, Asialoglycans, Gal/GalNAcβ1-4GlcNAcβ1-6Gal/GalNAc

Gal1-S 072-06341

078-06343 1mg

1mg×5

cDNA Source : Human

Target Glycan : branched LacNAc

Gal1, Stable Form 070-06401 50μg cDNA Source : Human

Target Glycan : branched LacNAc, Gal

Gal2 073-06371 1mg cDNA Source : Human

Target Glycan : GalNAcα1-3Gal (Blood Group A), branched LacNAc

Gal3C-S 079-06351

075-06353 1mg

1mg×5

cDNA Source : Human

Target Glycan : poly LacNAc

Gal3(Full-length) 077-06411 50μg cDNA Source : Human

Target Glycan : poly LacNAc

Gal7-S 076-06361

072-06363 1mg

1mg×5

cDNA Source : Human

Target Glycan : Galβ1-3GlcNAc

Gal9, Stable Form 074-06421 50μg cDNA Source : Human

Target Glycan : poly LacNAc, GalNAcα1-3Gal (Blood Group A)

LSL-N 127-06361

123-06363 1mg

1mg×5

cDNA Source : Laetiporus sulphureus

Target Glycan : LacNAc, poly LacNAc

MOA 137-18011

133-18013 1mg

1mg×5

cDNA Source : Marasmius oreades

Target Glycan : Galα1-3[Fucα1-2]Galβ1-4GlcNAc (Blood Group B)

Galα1-3Galβ1-4GlcNAc, Galα1-3Gal

PA-IL 167-26721

163-26723 1mg

1mg×5

cDNA Source : Pseudomonas aeruginosa

Target Glycan : Galα1-3(4)Gal

PPL 168-26751 0.5mg cDNA Source : Pleurocybella porrigens

Target Glycan : α/βGalNAc

SRL 199-17271

195-17273 1mg

1mg×5

cDNA Source : Sclerotium rolfsii

Target Glycan : Galβ1-3GalNAc (T antigen), Galβ1-3GlcNAc (Lacto-N-biose)

XCL 241-00971 1mg cDNA Source : Xerocomus chrysenteron

Target Glycan : Galβ1-3GalNAc/GlcNAc

Sialic Acid

ACG 011-25431

017-25433 1mg

1mg×5

cDNA Source : Agrocybe cylindracea

Target Glycan : α2-3 Sialic Acid

PSL1a 161-26741

167-26743 0.5mg

0.5mg×5

cDNA Source : Polyporus squamosus

Target Glycan : α2-6 Sialic Acid

Mannose

-Complex

BanLec 025-18661 0.5mg

cDNA Source : Musa acuminate

Target Glycan : α-Mannan, α-Glucan (Recognize internal α1-3 linked

glucosyl residues. Not recognize α-1-4, 1-6 linkage)

BC2L-A 026-18691 1mg cDNA Source : Burkholderia cenocepacia

Target Glycan : High-mannose

Calsepa 031-23831

037-23833 1mg

1mg×5

cDNA Source : Calystegia sepium

Target Glycan : High-mannose

GRFT 070-06381

076-06383 1mg

1mg×5

cDNA Source : Griffithia sp.

Target Glycan : High-mannose

Heltuba 086-10331 1mg cDNA Source : Helianthus tuberous

Target Glycan : Mannose

PALa 164-26731

160-26733 1mg

1mg×5

cDNA Source : Phlebodium aureum

Target Glycan : High-mannose

Orysata 159-03281

155-03283 1mg

1mg×5

cDNA Source : Oryza sative

Target Glycan : High-mannose

【Product Outline】

○Formulation:Liquid

○Source：Each lectin cDNA (synthetic) expressed in E. coli.

○Storage Condition：－20℃ and/or －80℃ After thawing, keep at 2-10℃. Avoid the freeze-thaw cycle.

16400開110

Description Code No. Package Size Glycan Specificity

Concanavalin A [Con A]

037-08771

033-08773

031-08774

100mg

250mg

1g

Man

Datura Lectin, Lyophilized [DSA] 047-23231 5mg Glu, GlcNAc

Dolichos Biflorus Lectin, Peroxidase Labeled [DBA] 047-23731 1mg GalNAc

Maackia Amurensis Lectin, Lyophilized [MAM] 139-10891 2mg Gal, GalNAc

Peanut Lectin, Lyophilized [PNA] 165-15031 5mg Gal, GalNAc

Phytohemagglutinin-E4, Lyophilized [PHA-E4] 164-15241 5mg Glc, GlcNAc

Phytohemagglutinin-L4 [PHA-L4] 168-15261 5mg Glc, GlcNAc

Phytohemagglutinin-P [PHA-P] 161-15251 50mg Glc, GlcNAc

Sambucus Sieboldiana Lectin, Lyophilized [SSA] 197-10371 5mg Siaα2-6Gal, Siaα2-6GalNAc

Soybean Lectin, Lyophilized 198-09931 5mg Gal, GalNAc

Wheat Germ Lectin [WGA] 126-02811 10mg Glc, GlcNAc

Description Code No. Package Size Clone No. Host Species Isotype Reference

Anti Siaα2-3, Monoclonal Antibody

recognition region: GM3, GM4, α2-3nLc4Cer

011-25171 200μL HYB4 Mouse IgG3･κ 1) Hidari, K. et al., BBRC, 436, 394

(2013).

Anti Keratan Sulfate, Monoclonal Antibody

recognition region: Keratan sulfate (lacking

oversulfated structure)

011-25811

017-25813

200μL

1mL R-10G Mouse IgG1

1) Kawabe, K. et al., Glycobiology, 23(3),

322 (2013).

Anti Sialyl-Tn Antigen, Monoclonal Antibody

recognition region: A cluster composed of Siaα2-

6GalNAc-Ser/Thr (Sialyl Tn),

Mucin-type glycoprotein

010-25881

016-25883

100μL

500μL MLS132 Mouse IgG3

1) Fukui, S. et al., Jpn. J. Cancer Res.,

79, 1119 (1988).

2) Numata, Y. et al., BBRC, 170(3), 981

(1990).

3) Nakada, H. et al., BBRC, 187(1), 217

(1992).

4) Nakada, H. et al., Proc. Natl. Acad.

Sci. USA, 90, 2495 (1993).

5) Tanaka, H. et al., Eur. J. Biochem.,

263, 27 (1999).

6) Akita, K. et al., Int. J. Gynecol.

Cancer, 22, 531 (2012).

Anti Tn Antigen, Monoclonal Antibody

recognition region: A cluster composed of

GalNAc-Ser/Thr (Tn antigen)

017-25891

013-25893

100μL

500μL MLS128 Mouse IgG3

Anti Sialyl-Lea Antigen, Monoclonal Antibody

recognition region: Siaα2-3Galβ1-3[Fucα1-4]GlcNAc

(Sialyl Lea), Siaα2-3Galβ1-3GlcNAc

010-25901

016-25903

100μL

500μL MSW113 Mouse IgG3

1) Kitagawa, H. et al., J. Biochem., 104,

817 (1988).

2) Kitagawa, H. et al., J. Biol. Chem.,

268(35), 26541 (1993).

3) Akita, K. et al., Int. J. Gynecol.

Cancer, 22, 531 (2012).

Antibodies to Carbohydrate Antigen

Storage Condition

： 2～10℃

： －20℃

： －80℃

TRA-1-81

R-10G

DAPI Merge Phase

DAPI Merge Phase

Detection of undifferentiated iPS cells using R-10G antibody

＜Detection condition＞ Primary Antibody : Upper Section : TRA-1-81 antibody as positive control Lower Section : Anti Keratan Sulfate antibody (clone No. R-10G) as test Secondary Antibody : FITC labeled secondary antibodies Cells: Human iPS cells 201B7 strain. Cultured on the feeder cells.

Bulk Supply is Available! Let’s contact with WAKO.