PathScan? Phospho-IGF-I Receptor β (Tyr1131) Sandwich ELISA Kit

• 产品编号：CST-7302C      品牌：CST       原厂货号：7302C
• 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-IGF-I Receptor β (Tyr1131) Sandwich ELISA Kit #7302用于检测内源性在1131位酪氨酸残基磷酸化的IGF-I受体β蛋白。用IGF-I处理MCF-7细胞后，IGF-I受体β蛋白在1131位酪氨酸残基磷酸化水平明显提高（图1）。而Western检测所示总的IGF-I受体β蛋白（磷酸化和未磷酸化的）没有改变（图1）。内部实验显示，该抗体主要识别相应物种的相关蛋白，但也可与其它物种的同源蛋白交互反应。PathScan® Phospho-IGF-I Receptor beta (Tyr1131) Sandwich ELISA Kit是一种固相三明治酶联免疫试验(ELISA)试剂盒，用于检测内源性在1131位酪氨酸残基磷酸化的IGF-I受体β蛋白。步骤如下：首先在微孔中结合有固相的兔抗磷酸化IGF-I（Tyr1131）抗体，然后将细胞裂解液于固相抗体中孵育，磷酸化IGF-I（Tyr1131）被固相抗体所捕获。经过洗脱后，非特异性结合的成分被洗脱下来，然后加入鼠源源磷酸化IGF-I（Tyr1131）检测抗体来检测被捕获的磷酸化IGF-I蛋白，然后再加入辣根过氧化物酶耦联的抗体来检测特异性结全的检测抗体。最后加入辣根过氧化物酶的底物TMB来进行显色。磷酸化IGF-I（Tyr1131）的浓度与显色的强度成正比，因此可以借此对磷酸化IGF-I（Tyr1131）定量。试剂盒中的抗体是定制的。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-IGF-I Receptor β (Tyr1131) Sandwich ELISA Kit #7302 detects endogenous levels of IGF-I Receptor β protein when phosphorylated at Tyr1131. A significant induction of phosphorylation of IGF-I receptor at Tyr1131 is detected in IGF-I- treated MCF-7 cells using PathScan® Phospho-IGF-I Receptor β (Tyr 1131) Sandwich ELISA Kit #7302 (Figure 1). The levels of total IGF-I receptor β (phospho and nonphospho) shown by Western analysis remain unchanged (Figure 1). 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-IGF-I Receptor beta (Tyr1131) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IGF-I receptor beta protein when phosphorylated at Tyr1131. A Phospho-IGF-I Receptor beta (Tyr1131) Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, phospho-IGF-I Receptor beta is captured by the coated antibody. Following extensive washing, an IGF-I Receptor Mouse Antibody is added to detect the captured phospho-IGF-I receptor protein. Anti-mouse IgG, HRP-linked Antibody #7076 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of IGF-I receptor protein phosphorylated at Tyr1131. 
 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).