Western Blot Antibody Customer Review for Anti-ALK Polyclonal Antibody (STJ91562)

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Anaplastic lymphoma kinase also known as ALK tyrosine kinase receptor or CD246 (cluster of differentiation 246) is an enzyme that in humans is encoded by the ALK gene. ALK plays an important role in the development of the brain and exerts its effects on specific neurons in the nervous system. Transduces signals from ligands at the cell surface, through specific activation of the mitogen-activated protein kinase (MAPK) pathway.The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. Anti-ALK - http://www.stjohnslabs.com/alk-antibody-p-91127?filter_name=STJ91562 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

Science

Result
“In H2228 and H3122 cell line, there were target bands detected by STJ91562. Anti-ALK antibody . Cost effectiveness,
specific binding, reasonable dilution requirements, low background “ - R. Wei Western University, Canada
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.
Lane No. Antigen
Loading
amount
Primary anti-
body
Primary anti-
body dilution
ratio
Secondary
antibody dilution
ratio
Target band KD
Visualiza-
tion time
1 H2228 10 ug
STJ91562.
ALK antibody
1:1000
1:5000 ~86KD 60S
2 H3122 10 ug 1:5000 ~117KD 60S
Cell Line: H2228, H3122
Method of validation: Western Blot
Primary Antibody: STJ91562 Anti -ALK Antibody
Secondary Antibody HRP goat anti-rabbit
Reference antibody: β-Tubulin
Dilution ratio: 1:1000
Protocol
Treatment of materials:
When cells were cultured to 70%-80% plating density, lysis buffer with protease inhibitors (Roche) and equal volume of
2X loading buffer were added for 10 min water bath at 100˚C.
E.g. Gel electrophoresis: 8% polyacrylamide gel, constant voltage 150 V, 60 min.
Transfer: 0.45 µm PVDF membrane, 1 piece of gel (semi-dry transfer), constant voltage 20 V 45 min.
Blocking: 1X TBST with 3% BSA for 1 hr.
Primary antibody probing: primary antibody was added to the solution of 1X TBST with 3% BSA, 4˚C for overnight.
Membrane wash: 1X TBST wash for 3 times.
Secondary antibody probing: secondary antibody was added to the solution of 1X TBST with 5% skimmed milk powder
with 1:5000 dilution ratio, for 1 hr.
Membrane wash: 1X TBST wash for 3 times.
Visualization: ECL visualization for 5 min (the visualization time was determined by actual result. It was ensured that
the visualization time was identical for the same antigen).
Antibody Customer Review:
STJ91562 Anti-ALK Antibody
Antibody Specificity:
Antibody Rating:
Figure: Lane1: H2228; Lane 2: H3122

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