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Custom Ordering Details: Product is assembled upon order to ensure maximum activity. United States: Please allow up to two weeks for your order to be processed and shipped. Outside of the United States: Please allow up to three weeks, depending on the country, for your order to be processed and shipped. When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

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REACTIVITY
Product Includes Volume Solution Color
IRS-1 Ab Coated Microwells 96 tests
P-IRS-1 (S302) Detection Ab 11 ml Green
Anti-rabbit IgG, HRP-linked Antibody 11 ml Red
TMB Substrate 7004 11 ml Colorless
STOP Solution 7002 11 ml Colorless
Sealing Tape 2 sheets
ELISA Wash Buffer (20X) 25 ml Colorless
Sample Diluent 25 ml Blue
Cell Lysis Buffer (10X) 9803 15 ml Yellowish

Product Description

The PathScan® Phospho-IRS-1 (Ser302) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IRS-1 when phosphorylated at Ser302. An IRS-1 Mouse Antibody* has been coated onto the microwells. After incubation with cell lysates, IRS-1 (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Phospho-IRS-1 (Ser302) Rabbit Detection Antibody* is added to detect phosphorylation of Ser302 on the captured IRS-1 protein. Anti-rabbit IgG, HRP-linked Antibody #7074* is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of IRS-1 phosphorylated at Ser302.

* Antibodies in kit are custom formulations specific to kit.


Specificity / Sensitivity

CST's PathScan® Phospho-IRS-1 (Ser302) Sandwich ELISA Kit #7283 detects endogenous levels of IRS-1 when phosphorylated at Ser302. As shown in Figure 1, a significant induction of IRS-1 phosphorylation at Ser302 can be detected in hSkMC cells following treatment with insulin using the Phospho-IRS-1 (Ser302) Sandwich ELISA Kit #7283. The level of total IRS-1 (phospho and nonphospho) remains unchanged as shown by Western analysis and by PathScan® Total IRS-1 Sandwich ELISA Kit #7328 (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.


Insulin receptor substrate 1 (IRS-1) is one of the major substrates of the insulin receptor kinase (1). IRS-1 contains multiple tyrosine phosphorylation motifs that serve as docking sites for SH2-domain containing proteins that mediate the metabolic and growth-promoting functions of insulin (2-4). IRS-1 also contains over 30 potential serine/threonine phosphorylation sites. Ser307 of IRS-1 is phosphorylated by JNK (5) and IKK (6) while Ser789 is phosphorylated by SIK-2, a member of the AMPK family (7). The PKC and mTOR pathways mediate phosphorylation of IRS-1 at Ser612 and Ser636/639, respectively (8,9). Phosphorylation of IRS-1 at Ser1101 is mediated by PKCθ and results in an inhibition of insulin signaling in the cell, suggesting a potential mechanism for insulin resistance in some models of obesity (10).


1.  Sun, X.J. et al. (1991) Nature 352, 73-7.

2.  Sun, X.J. et al. (1992) J Biol Chem 267, 22662-72.

3.  Myers, M.G. et al. (1993) Endocrinology 132, 1421-30.

4.  Wang, L.M. et al. (1993) Science 261, 1591-4.

5.  Rui, L. et al. (2001) J Clin Invest 107, 181-9.

6.  Horike, N. et al. (2003) J. Biol. Chem. 278, 18440-18447.

7.  Gao, Z. et al. (2002) J Biol Chem 277, 48115-21.

8.  Ozes, O.N. et al. (2001) Proc Natl Acad Sci U S A 98, 4640-5.

9.  De Fea, K. and Roth, R.A. (1997) Biochemistry 36, 12939-47.

10.  Li, Y. et al. (2004) J Biol Chem 279, 45304-7.


Entrez-Gene Id 3667
Swiss-Prot Acc. P35568


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a trademark of Cell Signaling Technology, Inc.