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3015
Insulin Receptor Substrate Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Insulin Receptor Substrate Antibody Sampler Kit #3015

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Insulin Receptor Substrate Antibody Sampler Kit: Image 1

Western blot analysis of MCF-7 cell extracts, unstimulated and insulin-stimulated (100 nM for 5 min), using IRS-1 Antibody #2382 (left) and Phospho-IRS-1 (Ser307) Antibody (right).

Insulin Receptor Substrate Antibody Sampler Kit: Image 2

Western blot analysis of cell extracts from CHO IR/IRS-1 cells, untreated or treated with insulin, using Phospho-IRS-1 (Ser612) (C15H5) Rabbit mAb (upper and middle) or IRS-1 Antibody #2382 (lower). The middle blot was treated with calf intestinal phosphatase (CIP) before antibody probing.

Insulin Receptor Substrate Antibody Sampler Kit: Image 3

Western blot analysis of extracts from CHO IR/4PS cells (transfected with insulin receptor and IRS-2) or CHO IR/IRS-1 cells (transfected with insulin receptor and IRS-1), showing no change in total protein levels with insulin stimulation (100 nM for 5 min), using IRS-2 Antibody (top) or IRS-1 Antibody #2382 (bottom).

Insulin Receptor Substrate Antibody Sampler Kit: Image 4

Western blot analysis of extracts from serum-starved C2C12 cells, untreated or insulin-treated (150 nM for 5 min.), using Phospho-IRS-1 (Ser318) (D51C3) Rabbit mAb (upper), or IRS-1 Antibody #3407 (lower).

Insulin Receptor Substrate Antibody Sampler Kit: Image 5

Western blot analysis of MCF7, C2C12 and RD cell lines using IRS-1 (D23G12) Rabbit mAb.

Insulin Receptor Substrate Antibody Sampler Kit: Image 6

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Insulin Receptor Substrate Antibody Sampler Kit: Image 7

Western blot analysis of extracts from CHO IR/IRS-1 cells (transfected with insulin receptor and IRS-1), untreated or insulin-treated (100 nM for 5 min), showing an increase in phospho-IRS-1 (Ser307) with insulin stimulation, using Phospho-IRS-1 (Ser307) Antibody.

To Purchase # 3015T
Product # Size Price
3015T
1 Kit  (5 x 20 µl) $ 391

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-IRS-1 (Ser307) Antibody 2381 20 µl
  • WB
H M R 180 Rabbit 
Phospho-IRS-1 (Ser612) (C15H5) Rabbit mAb 3203 20 µl
  • WB
H M R 180 Rabbit IgG
IRS-2 Antibody 4502 20 µl
  • WB
H M R 185 Rabbit 
Phospho-IRS-1 (Ser318) (D51C3) Rabbit mAb 5610 20 µl
  • WB
  • IP
H M 180 Rabbit IgG
IRS-1 (D23G12) Rabbit mAb 3407 20 µl
  • WB
  • IP
H M R Mk 180 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Insulin Receptor Substrate Antibody Sampler Kit provides an economical means to investigate IRS-1 and IRS-2 signaling and phosphorylation within the cell. The kit contains enough antibody to perform two western blots with each primary antibody.

Specificity / Sensitivity

Phospho-IRS-1 (Ser307) Antibody, Phospho-IRS-1 (Ser612) (C15H5) Rabbit mAb and Phospho-IRS-1 (Ser318) Antibody detect endogenous levels of IRS-1 only when phosphorylated at Ser307, Ser612 or Ser318, respectively. Note: Ser318 is the mouse residue; the corresponding human residue is Ser323. IRS-1 Antibody and IRS-2 Antibody detect endogenous levels of total IRS-1 or IRS-2, respectively. The Phospho-IRS-1 (Ser318) antibody cross reacts with a inducible nonspecific band at around 57 kDa.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy-terminal sequence of human IRS-2 and residues surrounding Ser307 of mouse IRS-1 (homologous to Ser312 of human IRS-1). Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser612 of mouse IRS-1, a synthetic peptide corresponding to the sequence surrounding Ser270 of human IRS-1 and a synthetic phosphopeptide corresponding to residues surrounding Ser318 of mouse IRS-1 protein.

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.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.