Products:Signal Transduction >> Protein Phosphorylation >> Tyrosine Kinases >> Receptor Tyrosine Kinases
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Testing dicount for ab5681to use in rat tissue. |
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i'm interested in detecting insulin receptor activation and have been looking at the literature. It is stablished that autophosphorylation of InsR,to undergo activation, takes place at tyr 1146, tyr 1150 and tyr1151. My question is ,why this phosphospecific Ab recognize phorylation at site 1158, 1162 and 1163? is it because the sequence is in different species? arr those sites known to be phosphorylated aswell, when binding of insulin to the receptor? thanks in advance for your time.
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We have just received the following information from the source of ab5681: The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of IR/IGF1R that contains tyrosines 1158, 1162 and 1163 of the human insulin receptor (IR) as numbered according to Ebina, et al. (1146, 1150 and 1151 according to Ullrich, et al.). The corresponding residues in the IGF1R are 1131, 1135 and 1136. The sequence is conserved in human, mouse and rat for both the IR and IGF1R.The two relevant papers are:Ebina, Y., et al. (1985) The human insulin receptor cDNA: the structural basis for hormone-activated transmembrane signalling. Cell 40(4):747-758.Ullrich, A., et al. (1985) Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. Nature 313(6005):756-761.When I look in NCBI, I see that there is a long and a short isoform of this protein. This is why we are listing 1158, 1162 and 1163 in the name (where these phospho sites in the long isoform) as well as 1146, 1150 and 1151 (for the short isoform) within the TDS. I hope this information helps, please do not hesitate to contact us again if you require further assistance, |
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Peptide Competition: Extracts prepared from CHO-T cells transfected with an insulin receptor containing vector and left unstimulated (1) or stimulated with insulin (2-8) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA TBST buffer overnight at 4°C and incubated with the ab5681 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphorylated peptide corresponding to the immunogen (3), a generic phosphotyrosine-containing peptide (4), the phosphopeptides corresponding to other IR/IGF1R sites (5-7) or, the phosphopeptide immunogen (8). After washing, membranes were incubated with goat F(ab’ 2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that only the phosphopeptide corresponding to IR/IGF1R [pYpYpY1158/1162/1163] completely blocks the antibody signal, thereby demonstrating the specificity of the antibody. The data also show the activation of the insulin receptor upon stimulation with insulin.
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