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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 um 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 um 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 15ug 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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