Phospho-ErbB-3 (Y1197) also known as phosphor receptor tyrosine-protein kinase erbB-3 is a member of the epidermal growth factor tyrosine kinase receptor family. It is encoded by the ERBB3 gene and is a membrane bound protein. The Phospho-ErbB-3 (Y1197) Polyclonal Antibody detects endogenous levels of ErbB-3 protein when phosphorylated at Y1197.

1. HOW DID OUR
ANTIBODY PERFORM?
ANTIBODY CUSTOMER REVIEW: Phospho-ErbB-3 (Y1197) Polyclonal Antibody
(STJ90642 )

2. What was used?
Primary antibody: STJ90642 Phospho-ErbB-3 (Y1197)
Provider: St John's Laboratory
Dilution ratio: 1:1000
Application: Western Blot
Materials for validation: MCF7 cell line
Click for product data sheet PDF

3. What Was The Protocol?
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 minutes at 95˚C.
Gel Electrophoresis 10% Polyacrylamide gel with a constant voltage of 160V for 60 minutes.
Transfer 0.45 µm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100V for 75 minutes.
Blocking 1X TBST with 5% BSA for 1 hour.
Primary Antibody Probing Primary antibody was added to the solution of 1X TBST with 5% BSA, and left at 4˚C overnight.
Membrane Wash Membrane was washed with 1X TBST 3 times.
Secondary Antibody Probing Secondary antibody was added to the solution of 1X TBST with 5% BSA with a 1:2500 dilution
ratio, for 1 hour.
Membrane Wash Membrane was washed with 1X TBST 3 times.
Visualization ECL visualization for 3 min (the visualization time was determined by actual result. It was
ensured that the visualization time was identical for the same antigen).

4. What was the protocol?
Gel electrophoresis information: 10% polyacrylamide gel, constant voltage of 160V for 60 minutes.
Transfer information: 0.45 µm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 100V for 75 minutes.
No. Antigen
Loading
amount
Primary
antibody
Primary
antibody
dilution ratio
Secondary
antibody
dilution ratio
Target band KD
Visualization
time
1 MCF7 15µg
STJ96177
Phospho-ErbB-3
(Y1197)
antibody
1:1000 1:2500 ~170KD 40S

5. What were the results?

6. What did the customer think?
Antibody Specificity:
Antibody RPPA Rating:
Testing results were provided independently by R. Bernardo at the Royal College of
Surgeons in Ireland. They stated “In MCF7 cell line, there were target bands detected by
Phospho-ErbB-3 (Y1197) STJ90642 Antibody.”

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