Technical Data
I8700-04
IRS-1, phosphorylated (Ser307) (Insulin Receptor Substrate 1)
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Technical Data |
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I8700-04 |
IRS-1, phosphorylated (Ser307) (Insulin Receptor Substrate 1) |
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Description: IRS-1 is one of the major substrates of insulin receptor kinase. IRS-1 protein contains a conserved pleckstrin homology (PH) domain located at its amino terminus, adjacent to a phospho-tyrosine binding (PTB) domain. IRS-1 contains multiple tyrosine phosphorylation motifs that serve as docking sites for SH2 domain-containing proteins (Grb2, p85, SHP-2, Fyn and others) and which mediate the metabolic and growth-promoting functions of insulin. IRS-1 also contains over 30 potential Ser/Thr phosphorylation sites for kinases, such as PKA, PKC and MAPK. Once phosphorylated by insulin receptor, IRS-1 recruits the downstream components that mediate the insulin signaling. Applications: Suitable for use in Western Blot. Other applications not tested. Recommended Dilution: Western Blot: 1:1000-1:2000 detects phospho-IRS-1 (Ser307) in RIPA lysates from serum starved HEK 293 cells treated with 5ug/ml anisomycin for 30 minutes, but not in untreated cells. Optimal dilutions to be determined by the researcher. Immunoblot Analysis: HEK 293 cell lysates untreated and anisomycin stimulated were resolved by electrophoresis, transferred to nitrocellulose and probed with antiphospho-IRS-1 (Ser307) (1:1000 dilution) or anti-IRS-1 (1:1000 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
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