| Insulin exerts its effects by binding to the insulin receptor, a disulfide-linked heterotetrameric protein comprised of two alpha subunits and two beta subunits arranged in the following configuration: beta-alpha-alpha-beta. The alpha subunits each contain insulin binding sites and are entirely extracellular in localization. The beta subunits each possess an extracellular domain, a single transmembrane domain, and a cytoplasmic tyrosine kinase domain. Binding of insulin to the alpha subunits induces a conformation change in the receptor which activates the kinase domain, stimulating tyrosine autophosphorylation of the receptor and tyrosine phosphorylation of at least five different insulin receptor substrates designated IRS-1-4, and Shc. This tyrosine phosphorylation produces docking sites for proteins bearing SH2 domains, such as PI3-K, Grb-2, Nck, and Crk. While tyrosine phosphorylation of insulin receptor substrates propagates signaling from insulin, serine and threonine phosphorylation by MAPK’s, PKA, and PKC may reduce insulin signaling. |
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| Catalog # | I8700-18A |
| Applications | Suitable for use in ELISA, Western Blot and Immunoprecipitation. Other applications not tested. |
| Recommended Dilution | ELISA: 0.1-1.0ug/ml |
| Immunoprecipation: 3-5ug/extract from 10e7 cells |
| Western Blot: 0.5-2.0ug/ml |
| Optimal dilutions to be determined by the researcher. |
| Recommended Positive Control | Serum-treated mouse fibroblast (3T3) cells. |
| Storage and Stability | May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and 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 |
| Host | Rabbit |
| Source | Human |
| Concentration | ~0.5mg/ml |
| Form | Supplied as a liquid in PBS, pH 7.2. No preservative added. Also available with azide. See I8700-18. |
| Purity | Purified from rabbit serum by epitope affinity chromatography. |
| Immunogen | Synthetic phosphopeptide derived from the human IRS-2 sequence, corresponding to the serine 731 phosphorylation site. This site corresponds to serine 133 in mouse and serine 233 in rat. |
| Specificity | Recognizes human IRS-2 when phosphorylated at serine 731. |
| Species Crossreactivity | 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. |
