PathScan? Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit

• 产品编号：CST-7258C      品牌：CST       原厂货号：7258C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258用于检测Tyr1150/1151位磷酸化的胰岛素受体。1图所示为CHO-IR/IRS-1细胞经胰岛素处理后，Tyr1150/1151磷酸化的胰岛素受体水平明显提高，所用抗体为Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258。但是Western检测所示胰岛素受体β（磷酸化和未磷酸化的）总水平没有变化。同时NIH/3T3和HepG2经胰岛素处理后，用该试剂盒也可检测到Tyr1150/1151位磷酸化的胰岛素受体，内部试验证明，该抗体用于检测相应物种的蛋白，但也可能与其它物种的同源蛋白交叉反应。赛信通的PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit是一种固相三明治酶连免疫吸收试验（ELISA）试剂盒，用于检测转染的Tyr1150/1151磷酸化胰岛素受体蛋白。Tyr1150/1151胰岛素受体β鼠单抗首先结合于小孔表面，将细胞裂解液与抗体孵育后，磷酸化和非磷酸化的胰岛素受体蛋白都被固相抗体捕获。充分洗脱后，加入 Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb，用于检测被捕获的Tyr1150/1151磷酸化胰岛素受体蛋白。然后加入抗兔IgG的辣根过氧化物酶连抗体来识别特异结合的检测抗体。最后加入辣根过氧化物酶的底物，TMB进行显色。显色的吸光度与Tyr1150/1151磷酸化胰岛素受体蛋白的量成正比。试剂盒中的抗体是专门定做的。一型胰岛素样生长因子受体(IGF-IR)是一种跨膜的酪氨酸激酶，在许多新生及成熟的组织细胞中都广泛表达(1-3)。IGF-I和IGF-II配体结合到受体上去，受体就发生自我磷酸化。在激酶结构域，有三个酪氨酸残基(Tyr1131, Tyr1135,Tyr1136)是最先发生自身磷酸化的位点(4)。这三个残基的磷酸化对激酶的激活是必需的(5,6)。胰岛素受体(IRs)跟胰岛素样生长因子受体有着相似的结构和功能，在它的激酶活化环上同样有三个酪氨酸簇(Tyr1146/1150/1151)。胰岛素受体酪氨酸残基的磷酸化是细胞接受胰岛素刺激后最先发生的生化反应(7)。自身磷酸化最先在Tyr1146和Tyr1150(或Tyr1151)发生，但激酶的激活需要三者同时磷酸化(8)。

1. Adams, T.E. et al. (2000) Cell Mol Life Sci 57, 1050-93.
2. Baserga, R. (2000) Oncogene 19, 5574-81.
3. Scheidegger, K.J. et al. (2000) J Biol Chem 275, 38921-8.
4. Hernández-Sánchez, C. et al. (1995) J Biol Chem 270, 29176-81.
5. Lopaczynski, W. et al. (2000) Biochem Biophys Res Commun 279, 955-60.
6. Baserga, R. (1999) Exp Cell Res 253, 1-6.
7. White, M.F. et al. (1985) J Biol Chem 260, 9470-8.
8. White, M.F. et al. (1988) J Biol Chem 263, 2969-80.

原厂资料：

Specificity / Sensitivity

CST''s PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258 detects phospho-insulin receptor (Tyr1150/1151) protein. As shown in Figure 1, using Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258, a significant induction of phospho-insulin receptor (Tyr1150/1151) is detected in CHO-IR/IRS-1 cells treated with insulin. The levels of total insulin receptor β (phospho and nonphospho) shown by Western analysis remain unchanged. Endogenous phospho-insulin receptor β (Tyr1150/1151) in either NIH/3T3 or HepG2 cells treated with insulin is also detected by Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit #7258. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

CST''s PathScan® Phospho-Insulin Receptor β (Tyr1150/1151) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected phospho-insulin receptor (Tyr1150/1151) protein. An Insulin Receptor β Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-insulin receptor proteins are captured by the coated antibody. Following extensive washing, Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7) Rabbit mAb is added to detect the captured phospho-insulin receptor (Tyr1150/1151) protein. Anti-rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of phospho-insulin receptor β (Tyr1150/1151) protein. 
 * Antibodies in kit are custom formulations specific to kit.

Background

Type I insulin-like growth factor receptor (IGF-IR) is a transmembrane receptor tyrosine kinase that is widely expressed in many cell lines and cell types within fetal and postnatal tissues (1-3). Receptor autophosphorylation follows binding of the IGF-I and IGF-II ligands. Three tyrosine residues within the kinase domain (Tyr1131, Tyr1135, and Tyr1136) are the earliest major autophosphorylation sites (4). Phosphorylation of these three tyrosine residues is necessary for kinase activation (5,6). Insulin receptors (IRs) share significant structural and functional similarity with IGF-I receptors, including the presence of an equivalent tyrosine cluster (Tyr1146/1150/1151) within the kinase domain activation loop. Tyrosine autophosphorylation of IRs is one of the earliest cellular responses to insulin stimulation (7). Autophosphorylation begins with phosphorylation of Tyr1146 and either Tyr1150 or Tyr1151, while full kinase activation requires triple tyrosine phosphorylation (8).

1. Adams, T.E. et al. (2000) Cell Mol Life Sci 57, 1050-93.
2. Baserga, R. (2000) Oncogene 19, 5574-81.
3. Scheidegger, K.J. et al. (2000) J Biol Chem 275, 38921-8.
4. Hernández-Sánchez, C. et al. (1995) J Biol Chem 270, 29176-81.
5. Lopaczynski, W. et al. (2000) Biochem Biophys Res Commun 279, 955-60.
6. Baserga, R. (1999) Exp Cell Res 253, 1-6.
7. White, M.F. et al. (1985) J Biol Chem 260, 9470-8.
8. White, M.F. et al. (1988) J Biol Chem 263, 2969-80.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).