Product Pathways - Metabolism
IRS-1 Inhibition Antibody Sampler Kit #12879
|12879S||1 Kit (7 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Phospho-IRS-1 (Ser302) (34C7) Rabbit mAb #2491||40 µl||W||H, M||180||Rabbit IgG|
|Phospho-IRS-1 (Ser307) Antibody #2381||40 µl||W, IP||H, M, R||180||Rabbit|
|Phospho-IRS-1 (Ser318) (D51C3) Rabbit mAb #5610||40 µl||W, IP||H, M||R||180||Rabbit IgG|
|Phospho-IRS-1 (Ser612) (C15H5) Rabbit mAb #3203||40 µl||W||H, M, R||180||Rabbit IgG|
|Phospho-IRS-1 (Ser636/639) Antibody #2388||40 µl||W||H, M, R||180||Rabbit|
|Phospho-IRS-1 (Ser1101) Antibody #2385||40 µl||W, IP||H, M, R||180||Rabbit|
|IRS-1 (D23G12) Rabbit mAb #3407||40 µl||W, IP||H, M, R, Mk||180||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey
Western blot analysis of extracts from CHO IR/IRS-1 cells (transfected with insulin receptor and IRS-1), untreated (-) or insulin-treated (100 nM, 5 min; +), showing an increase in phospho-IRS-1 (Ser307) with insulin stimulation, using Phospho-IRS-1 (Ser307) Antibody #2381.
Western blot analysis of IRS-1 Antibody #2382 immunoprecipitated pellets from differentiated C2C12 cells, unstimulated (-) or stimulated with insulin (100 nM, 15 min; +) or PMA (200 mM, 30 min; +), using Phospho-IRS-1 (Ser1101) Antibody #2385 (upper) or IRS-1 Antibody #2382 (lower).
Western blot analysis of extracts from C2C12 cells, untreated (-) or insulin-treated (100 nM, 5 min; +), using Phospho-IRS-1 (Ser636/639) Antibody #2388.
Western blot analysis of insulin treated (100 nM, 5 min) MCF7 and C2C12 cells using Phospho-IRS-1 (Ser302) (34C7) Rabbit mAb #2491.
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 #3203 (upper and middle) or IRS-1 Antibody #2382 (lower). The middle blot was treated with calf intestinal phosphatase (CIP) before antibody probing.
Western blot analysis of MCF7, C2C12 and RD cell lines using IRS-1 (D23G12) Rabbit mAb #3407.
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 four 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).
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).
- Sun, X.J. et al. (1991) Nature 352, 73-7.
- Sun, X.J. et al. (1992) J Biol Chem 267, 22662-72.
- Myers, M.G. et al. (1993) Endocrinology 132, 1421-30.
- Wang, L.M. et al. (1993) Science 261, 1591-4.
- Cao, Y. et al. (2006) J Biol Chem 281, 40242-51.
- Nakamori, Y. et al. (2006) J Cell Biol 173, 665-71.
- Rui, L. et al. (2001) J Clin Invest 107, 181-9.
- Gao, Z. et al. (2002) J Biol Chem 277, 48115-21.
- Greene, M.W. et al. (2004) Biochem J 378, 105-16.
- Weigert, C. et al. (2006) J Biol Chem 281, 7060-7.
- del Rincón, S.V. et al. (2004) Oncogene 23, 9269-79.
- Moeschel, K. et al. (2004) J Biol Chem 279, 25157-63.
- Ozes, O.N. et al. (2001) Proc Natl Acad Sci U S A 98, 4640-5.
- De Fea, K. and Roth, R.A. (1997) Biochemistry 36, 12939-47.
- Li, Y. et al. (2004) J Biol Chem 279, 45304-7.
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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.