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Independent Antibody Validation Reviews
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What is Independent Validation?
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These up-to-date results are independently compiled and offer
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ANTIBODY VALIDATION REVIEWS
HA-tag Polyclonal Antibody (STJ90107)
Antibody: STJ90107. HA-tag Polyclonal
Antibody
Method for validation: Western Blot
Materials for validation:Hela cell line,
Jurkat cell line
Protocol: When cells were cultured to
70%-80% plating density, lysis buffer
with protease inhibitors (Sigma) and
equal volume of 2X loading buffer
were added for 10 min water bath at
100˚C.
Result of validation: target band was
found in the material cell lines.
Cdc2 Polyclonal Antibody (STJ92154)
Antibody: STJ92154. Cdc2
Polyclonal Antibody
Method for validation: Western
Blot
Materials for validation:Hela cell
line, Jurkat cell line
Protocol: When cells were
cultured to 70%-80% plating
density, lysis buffer with protease
inhibitors (Sigma) and equal
volume of 2X loading buffer were
added for 10 min water bath at
100˚C.
Result of validation: target band
was found in the material cell
lines.
FAAH Antibody (STJ23602)
Antibody:STJ23602. FAAH Antibody
Method for validation: Immunofluorescence
Materials for validation: Dorsal root ganglia
stained with FAAH antibody
Protocol:The sections processed for
immunofluorescence were studied with a
fluorescence microscope (DM RXA2) with
confocal scanner (TCS SL) (Leica
Microsystems GmbH, Manheim, Germany)
equipped with the following lasers: Ar 488,
He-Ne 543, He-Ne 633.
Result of validation: We completed
evaluation of FAAH antibody. It worked
really well, so we decided to purchase a vial
from St John’s Laboratory.
Actin-beta antibody (STJ91464)
Gel electrophoresis information：10% polyacrylamide gel, constant voltage 160 V, 60 min.
Transfer information：0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage
100 V 60 min.
No. Antigen Loading
amount
Primary
antibody
Primary
antibody
dilution
ratio
Secondary
antibody
dilution
ratio
Target band
KD
Visualiz
ation
time
1 Hela
lysate 10 ug
STJ9146
4.
Actin-
beta
antibody
1:1000
1:10000 ~50KD 20S
2 Jurkat
lysate 10 ug 1:10000 ~50KD 20S
Result of validation:
In Hela and Jurkat cell line,
there were target bands
detected by STJ91464.Actin-
beta antibody (Figure: Lane1:
Hela; Lane 2: Jurkat).
Synuclein-α antibody (STJ95862)
Antibody: STJ95862. Synuclein-α
antibody
Method for validation: IHC
Materials for validation:rat substantia
nigra, fixed frozen tissue
Antibody performance:clear staining of
endogenous alpha-synuclein protein
(on rat microglial cells). Staining is red in
image.
Result of validation:Clear staining of
endogenous alpha-synuclein protein.
Staining is red (OX42 – marker for
microglia) in image blue in cell nuclei
with DAPI.
EGFR antibody (STJ40568)
Antibody: STJ40568. EGFR antibody
Method for validation: Western Blot
Materials for validation:loading control:
β-actin, secondary antibody: Sheep
anti-mouse IgG-HRP
Antibody performance:Lysates were
prepared by Kinexus Bioinformatics
Corporation following standard
protocols and quality controlled for
protein integrity on a regular basis.
Result of validation:A strong band was
observed in the positive control at the
expected size (~170 kDa) that is not
observed in the negative control.
Antibody: STJ91464.Actin-beta antibody
Method for validation: Western Blot
Materials for validation:Hela cell line, HEPG2 cell line
Treatment of materials: When cells were cultured to 70%-80% plating
density, lysis buffer with protease inhibitors (Sigma) and equal volume
of 2X loading buffer were added for 10 min water bath at 100˚C.
mCherry Tag antibody (STJ34373)
Antibody: STJ34373. mCherry Tag
antibody
Method for
validation:Immunofluorescence
Materials for validation: Positive
control: A549 cells transfected with
mCherry vector pLV-mCherry stained
with mCherry antibody
Negative control: Untransfected A549
cells stained with mCherry antibody
Secondary Only control: A549 cells
transfected with mCherry vector and
stained with Goat anti-mouse
IgG+IgM Antibody (DyLight 488) only.
Isotype control: A549 cells transfected
with mCherry vector and stained with
isotype control mouse IgG1 (Cusabio).
Figure 1: A549 cells
transfected with mCherry
vector stained with mCherry
antibody (green). mCherry
itself also gives fluorescence
signal (red). Bottom-right
panel shows merged image
from blue, green, and red
channels.
PIWIL2 antibody (STJ25008)
Antibody: STJ25008.PIWIL2
antibody
Method for validation: Western
Blot
Materials for validation:rat testis
Treatment of materials:Ab gave
conclusive results with bands of
the correct size
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