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12879
IRS-1 Inhibition Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

IRS-1 Inhibition Antibody Sampler Kit #12879

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IRS-1 Inhibition Antibody Sampler Kit: Image 1

Western blot analysis of insulin treated (100 nM, 5 min) MCF7 and C2C12 cells using Phospho-IRS-1 (Ser302) (34C7) Rabbit mAb.

IRS-1 Inhibition Antibody Sampler Kit: Image 2

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).

IRS-1 Inhibition Antibody Sampler Kit: Image 3

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).

IRS-1 Inhibition Antibody Sampler Kit: Image 4

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.

IRS-1 Inhibition Antibody Sampler Kit: Image 5

Western blot analysis of extracts from C2C12 cells, untreated or insulin-treated (100 nM, 5 min) using, Phospho-IRS-1 (Ser636/639) Antibody.

IRS-1 Inhibition Antibody Sampler Kit: Image 6

Immunprecipitation of Phospho-IRS-1 from insulin treated MCF7 cell extracts using Phospho-IRS-1 (Ser1101) antibody (Lane 1). Lane 2: No antibody control. Lane 3: Input control.

IRS-1 Inhibition Antibody Sampler Kit: Image 7

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

IRS-1 Inhibition Antibody Sampler Kit: Image 8

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.

IRS-1 Inhibition Antibody Sampler Kit: Image 9

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.

IRS-1 Inhibition Antibody Sampler Kit: Image 10

Western blot analysis of IRS-1 (#2382) antibody-immunoprecipitated pellets from differentiated C2C12 cells, unstimulated, insulin-stimulated (100 nM for 15 min) or PMA-stimulated (200 mM for 30 min), using Phospho-IRS-1 (Ser1101) Antibody (upper) or IRS-1 Antibody #2382 (lower).

To Purchase # 12879T
Product # Size Price
12879T
1 Kit  (7 x 20 µl) $ 500

Product Description

The IRS-1 Inhibition Antibody Sampler Kit provides an economical means to evaluate insulin signaling negative feedback loops via phosphorylation of various IRS-1 serine residues. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each activation state antibody recognizes the phosphorylated form of its target. All target residues are based on the sequence for mouse IRS-1, except Ser636/339 and Ser1101, which are based on the sequence for human IRS-1. IRS-1 (D23G12) Rabbit mAb recognizes total IRS-1 protein independent of its phosphorylation state.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding mouse Ser307 (human Ser312), human Ser636/639 (mouse Ser632/635), and human Ser1101 (mouse Ser1097) of IRS-1. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding mouse Ser302 (human Ser307), mouse Ser318 (human Ser323), and mouse Ser612 (human 616) of IRS-1. IRS-1 (D23G12) rabbit monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence surrounding Ser270 of human IRS-1 (mouse 1097).

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, many of which are related to negative feedback loops activated during insulin signaling. Ser302 (human Ser307) of IRS-1 is regulated by FOX01 (5), IKKγ, and MYO1C (6). Ser307 (human Ser312) of IRS-1 is phosphorylated by JNK (7) and IKK (8). PKC phosphorylates mouse IRS-1 at Ser318 (human Ser323) by insulin receptor activation or by other stimulation such as TPA, IL-6, and retinoic acid treatment (9-12). The PKC and mTOR pathways mediate phosphorylation of IRS-1 at Ser612 (human Ser616) and Ser632/635 (human Ser636/639), respectively (13,14). Phosphorylation of IRS-1 at Ser1097 (human 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 (15).

  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. Cao, Y. et al. (2006) J Biol Chem 281, 40242-51.
  6. Nakamori, Y. et al. (2006) J Cell Biol 173, 665-71.
  7. Rui, L. et al. (1997) J. Clin. Invest. 107, 181-189.
  8. Gao, Z. et al. (2002) J. Biol. Chem. 277, 48115-48121.
  9. Greene, M.W. et al. (2004) Biochem J 378, 105-16.
  10. Weigert, C. et al. (2006) J Biol Chem 281, 7060-7.
  11. del Rincón SV et al. (2004) Oncogene 23, 9269–79
  12. Moeschel, K. et al. (2004) J Biol Chem 279, 25157-63.
  13. Ozes, O.N. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 4640-4645.
  14. De Fea, K. and Ruth, R.A. (1997) Biochemistry 36, 12939-12947.
  15. 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.