Revision 1

#7288Store at 4C and -20C

 

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Species Cross Reactivity

UniProt ID:

#P35568

Entrez-Gene Id:

#3667

Cell Signaling Technology

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Volume Cap Color Storage Temp

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Description

CST's PathScan® Phospho-IRS-1 (Ser307) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-IRS-1 (Ser307) Sandwich ELISA Kit #7287. 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 (Ser307) 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 Ser307.
*Antibodies in this kit are custom formulations specific to the kit.

Reagents not supplied

Phosphate Buffered Saline (PBS-20X) #9808 Phosphate Buffered Saline with Tween-20 (PBST-20X) #9809 Cell Lysis Buffer (10X) #9803 TMB Substrate #7004 STOP Solution #7002 Blocking Buffer: 1X PBS/0.05% Tween-20, 1% BSA 96 Well Microplates** Microplate Reader ** Antibody Pairs have been validated on Corning® 96 Well Clear Polystyrene High Bind Stripwell™ Microplates (#2592) and Corning® 96 Well EIA/RIA Easy Wash™ Clear Flat Bottom Polystyrene High Bind Microplates (#3369).

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).

  1. Sun, X.J. et al. (1991) Nature 352, 73-77.
  2. Sun, X.J. et al. (1992) J. Biol. Chem. 267, 22662-22672.
  3. Myers Jr., M.G. et al. (1993) Endocrinology 132, 1421-1430.
  4. Wang, L.M. et al. (1993) Science 261, 1591-1594.
  5. Rui, L. et al. (1997) J. Clin. Invest. 107, 181-189.
  6. Gao, Z. et al. (2002) J. Biol. Chem. 277, 48115-48121.
  7. Horike, N. et al. (2003) J. Biol. Chem. 278, 18440-18447.
  8. Ozes, O.N. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 4640-4645.
  9. De Fea, K. and Ruth, R.A. (1997) Biochemistry 36, 12939-12947.
  10. Li, Y. et al. (2004) J. Biol. Chem. 279, 45304-45307.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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    Revision 1
    #7288

    PathScan® Phospho-IRS-1 (Ser307) Sandwich ELISA Antibody Pair

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