Independent Antibody Validation Reviews
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“Our independent validation reviews offer
scientists the opportunity to test our quality
and ensure reproducibility”.
What is Independent Validation?
Our aim is to provide an open platform that offers scientist’s from
academia and industry insights into how our products have
performed for the end-user.
These up-to-date results are independently compiled and offer
data about new applications, protocols, dilutions conditions and
images of our products in real-experimental conditions.
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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