Product Pathways - PathScan ELISA
PathScan® Phospho-IRS-1 (Ser302) Sandwich ELISA Antibody Pair #7284
| Kit Includes | Volume | Cap Color |
|---|---|---|
| IRS-1 Capture Ab (100X) | 0.4 ml | Pink |
| P-IRS-1 (S302) Detection Ab (100X) | 0.4 ml | Blue |
| Anti-rabbit IgG, HRP-linked Antibody (1000X) | 0.04 ml | Red |
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
Species Cross-Reactivity
H
Reactivity Key:
H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Important Ordering Details
Product is assembled upon order to ensure maximum activity. Domestic: Please allow up to two weeks for your order to be processed and shipped. International: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.
Protocols
- 7284:
- ELISA-P
Description
CST's PathScan® Phospho-IRS-1 (Ser302) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-IRS-1 (Ser302) Sandwich ELISA Kit #7283. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The IRS-1 Mouse Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysate is added followed by a Phospho-IRS-1 (Ser302) Rabbit Detection Antibody and an HRP-conjugated Anti-Rabbit IgG Antibody. HRP substrate (TMB) is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of IRS-1 phosphorylated at Ser302.*Antibodies in this kit are custom formulations specific to the kit.
Specificity / Sensitivity
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Sandwich ELISA
The relationship between the protein concentration of the lysate from untreated and insulin-treated hSkMC cells and the absorbance at 450 nm using PathScan® Phospho-IRS-1 (Ser302) Sandwich ELISA Antibody Pair #7284 is shown. After overnight starvation, hSkMC cells were treated with insulin (100 nM) for 15 minutes at 37ÂșC and then lysed.
Background
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).
- Sun, X.J. et al. (1991) Nature 352, 73-77.
- Sun, X.J. et al. (1992) J. Biol. Chem. 267, 22662-22672.
- Myers Jr., M.G. et al. (1993) Endocrinology 132, 1421-1430.
- Wang, L.M. et al. (1993) Science 261, 1591-1594.
- Rui, L. et al. (1997) J. Clin. Invest. 107, 181-189.
- Gao, Z. et al. (2002) J. Biol. Chem. 277, 48115-48121.
- Horike, N. et al. (2003) J. Biol. Chem. 278, 18440-18447.
- Ozes, O.N. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 4640-4645.
- De Fea, K. and Ruth, R.A. (1997) Biochemistry 36, 12939-12947.
- Li, Y. et al. (2004) J. Biol. Chem. 279, 45304-45307.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 7283 PathScan® Phospho-IRS-1 (Ser302) Sandwich ELISA Kit
- 7328 PathScan® Total IRS-1 Sandwich ELISA Kit
- 2382 IRS-1 Antibody
- 2390 IRS-1 (59G8) Rabbit mAb
- 3194 IRS-1 (L3D12) Mouse mAb
- 2491 Phospho-IRS-1 (Ser302) (34C7) Rabbit mAb
- 2384 Phospho-IRS-1 (Ser302) Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7004 TMB Substrate
- 7002 STOP Solution
- 9998 BSA
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
Rabbit Monoclonals are produced under license (granting certain rights, including those under U.S. Patent No. 5,675,063) from Epitomics, Inc.
For Research Use Only. Not For Use In Diagnostic Procedures.