Cell Signaling Technology

Product Pathways - Metabolism

Insulin Receptor Substrate Antibody Sampler Kit #3015

Insulin receptor substrate 1   insulin receptor substrate-1   insulin-receptor   insulin-receptor-substrate   IR   IRS   IRS 1   IRS-1   IRS1   rtkscaffold  

REACTIVITY

No. Size Price
3015T 1 Kit ( 5 x 20 µl ) ¥4,653.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
Phospho-IRS-1 (Ser307) Antibody #2381 20 µl W,IP, H,M,R, 180 Rabbit
Phospho-IRS-1 (Ser612) (C15H5) Rabbit mAb #3203 20 µl W, H,M,R, 180 Rabbit IgG
IRS-1 (D23G12) Rabbit mAb #3407 20 µl W,IP, H,M,R,Mk, 180 Rabbit IgG
IRS-2 Antibody #4502 20 µl W, H,M,R, 185 Rabbit
Phospho-IRS-1 (Ser318) (D51C3) Rabbit mAb #5610 20 µl W,IP, H,M, R, 180 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl W, Goat

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.

Phospho-IRS-1 (Ser307)抗体、Phospho-IRS-1 (Ser612) (C15H5)兔单抗和Phospho-IRS-1 (Ser318)抗体分别识别内源性的Ser307、Ser612或者Ser318磷酸化的IRS-1蛋白水平。注意:Ser318是小鼠大氨基酸残基,对应的人类氨基酸残基为Ser323。·IRS-1抗体和IRS-2抗体分别识别内源性的IRS-1或IRS-2总蛋白水平。Phospho-IRS-1 (Ser318)抗体也可以识别一条诱导的非特异的57kDa条带。

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.

该多克隆抗体用与人类IRS-2中羧基末端的氨基酸序列和小鼠IRS-1的Ser307(与人IRS-1 Ser312同源)附近的氨基酸序列对应的人工合成肽段免疫动物制成。该抗体使用蛋白A和肽亲和层析纯化而得。该单克隆抗体用与小鼠IRS-1中Ser612位点、人类IRS-1中Ser270位点或小鼠IRS-1中Ser318附近的氨基酸序列对应的人工合成肽段免疫动物制成。

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 four western blots with each primary antibody.

Insulin Receptor Substrate Antibody Sampler Kit提供了一个经济的手段来研究IRS-1和IRS-2信号通路及其在细胞内的磷酸化情况。该试剂盒包含足量的每个一抗进行四次Western blots的抗体。

Western Blotting

Western Blotting

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.

IP

IP

Immunoprecipitation of IRS-1 from differentiated C2C12 cell extracts, untreated or insulin-treated for the indicated times, using IRS-1 Antibody #2382, followed by Western blot with either Phospho-IRS-1 (Ser307) Antibody, or IRS-1 Antibody #2382.

Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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.

Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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.

Western Blotting

Western Blotting

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 #4502 (upper) or IRS-1 Antibody #2382 (lower).

对CHO IR/4PS细胞(转染胰岛素受体和IRS-2)或CHO IR/IRS-1细胞(转染胰岛素受体和IRS-1),使用 IRS-2 Antibody #4502(上图)或IRS-1 Antibody #2382(下图)进行Western blot分析,显示100nM胰岛素刺激5分钟总的蛋白水平没有变化。

Western Blotting

Western Blotting

Western blot analysis of extracts from MCF-7 cells, unstimulated or insulin-stimulated (100 nM for 5 min), using IRS-1 Antibody #2382 (left) and Phospho-IRS-1 (Ser307) Antibody #2381 (right).

对MCF-7细胞(未处理或150nM胰岛素处理5分钟),使用IRS-1 Antibody #2382(左)或Phospho-IRS-1 (Ser307) Antibody #2381(右)进行Western blot分析。

Western Blotting

Western Blotting

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

Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

Western blot analysis of MCF7, C2C12 and RD cell lines using IRS-1 (D23G12) Rabbit mAb #3407.

对MCF7、C2C12和RD细胞系使用IRS-1 (D23G12) Rabbit mAb兔单抗 #3407进行Western blot分析。

Western Blotting

Western Blotting

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 #5610 (upper), or IRS-1 Antibody #3407 (lower).

对血清饥饿的C2C12细胞(未处理或150nM胰岛素处理5分钟),使用Phospho-IRS-1 (Ser318) (D51C3) Rabbit mAb兔单抗 #5610 (upper)(上图)或IRS-1 Antibody #3407(下图)进行Western blot分析。

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).

胰岛素受体底物1(IRS-1)是胰岛素受体激酶的主要底物之一(1)。IRS-1包括多个酪氨酸磷酸化模体,作为与包含SH2结构域的蛋白质的接合位点,介导胰岛素的代谢和生长促进功能(2-4)。IRS-1也包含超过30个潜在的丝氨酸/苏氨酸磷酸化位点。JNK(5)和IKK(6)对IRS-1 Ser307位点进行磷酸化,SIK2(AMPK家族成员之一)对Ser789进行磷酸化(7)。PKC和mTOR途径分别介导IRS-1的Ser612和Ser636/639磷酸化(8,9)。IRS-1在Ser1101的磷酸化由PKCθ介导,可以导致细胞内胰岛素信号通路的抑制,这暗示其在某些肥胖模型中的胰岛素抵抗机制中发挥作用(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.

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For Research Use Only. Not For Use In Diagnostic Procedures.

U.S. Patent No. 5,675,063.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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