| 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 growthpromoting 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. |
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| Catalog # | I8700 |
| Applications | Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested. |
| Recommended Dilution | Western Blot: 0.5-2ug/ml detects IRS-1 in RIPA lysates from 3T3/A31 mouse fibroblasts. 3T3/A31 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with I8700 (0.5ug/ml). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. Previous lots detected IRS-1 in RIPA lysates of rat L6 myoblasts. In cells that contain high levels of IRS-1, this lot may work optimally at 0.1-0.5ug/ml. |
| Immunoprecipitation: 4ug immunoprecipitates IRS-1 from 500ug of 3T3/A31 RIPA lysate |
| Optimal dilutions to be determined by the researcher. |
| Positive Control | Fibroblast Cell, 3T3/A31, L6 myoblasts. |
| Recommended Secondary Antibodies |
| I1904-39: IgG, X-Adsorbed (HRP) Pab Gt xRb |
| I1904-40A: IgG, H&L, X-Adsorbed (HRP) Pab Gt xRb |
| I1904-46Q: IgG, H&L (HRP) Pab Gt xRb |
| 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. |
| CAS Number | n/a |
| Clone Type | Polyclonal |
| Isotype | IgG |
| Host | Rabbit |
| Source | Rat |
| Concentration | ~0.25mg/ml |
| Form | Supplied as a liquid in 0.2M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.1mM EDTA, 5mg/ml BSA, 0.05% sodium azide. |
| Purity | Purified by immunoaffinity chromatography. |
| Immunogen | Synthetic peptide corresponding to 14 amino acid ([C]YASINFQKQPEDRQ) with in the carboxy terminal of rat liver IRS-1. |
| Specificity | Recognizes IRS-1 at 160-185kD. Species Crossreactivity: Human, mouse and rat. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
