PathScan? Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit

• 产品编号：CST-7082C      品牌：CST       原厂货号：7082C
• 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 (panTyr) Sandwich ELISA Kit #7082用于检测转染的人酪氨酸残基磷酸化的胰岛素受体β蛋白，如1图所示。图2显示试剂盒的灵敏性。该试剂盒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-Insulin Receptor β (panTyr) Sandwich ELISA Kit是一种固相三明治酶联免疫试验(ELISA)试剂盒，用于检测转染的人酪氨酸残基磷酸化的胰岛素受体β蛋白。步骤如下：首先在微孔中结合有固相的胰岛素受体β蛋白抗体，然后将细胞裂解液于固相抗体中孵育，胰岛素受体β蛋白(磷酸化与非磷酸化的)蛋白被固相抗体所捕获。经过洗脱后，非特异性结合的成分被洗脱下来，然后加入磷酸化酪氨酸鼠单抗检测抗体来检测被捕获酪氨酸残基磷酸化的胰岛素受体β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。酪氨酸残基磷酸化的胰岛素受体β蛋白的浓度与显色的强度成正比，因此可以借此定量。胰岛素受体（INSR）是一种跨膜酪氨酸激酶。受体分子由二硫键相连的异二聚体组成。α亚基分子量为135 kDa，位于胞外段。β亚基分子量为95，包括一个胞外段，单次跨膜段和胞内酪氨酸激酶结构域（1）。胰岛素与受体结合后，会使受体自磷酸化和酪氨酸激酶活性激活。INSR可以催化一些分子如IRS, Gab1, Shc, and Cbl的酪氨酸残基磷酸化，这些分子可以进一步激活下游的MAPK, PI3K, and TC10信号通路。这最终会导致葡萄糖摄入增加和诸如细胞生长之类的代谢反应（2,3）。INSR跟其它的受体酪氨酸激酶一样，有着肽类底物的特异性，对在-1和-4位点有酸性残基和+1\+3位点的疏水残基有着强烈的偏好(4)。

1. Yip, C.C. and Ottensmeyer, P. (2003) J. Biol. Chem. 278, 27329-27332.
2. Saltiel, A.R. and Pessin, J.E. (2002) Trends Cell Biol. 12, 65-71.
3. Zick, Y. (2001) Trends Cell Biol. 11, 437-441.
4. Songyang, Z. and Cantley, L.C. (1995) TIBS 20, 470-475.

原厂资料：

Specificity / Sensitivity

PathScan® Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit #7082 detects transfected levels of human Insulin Receptor β protein phosphorylated at Tyrosine, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The PathScan® Phospho-Insulin Receptor β (panTyr) Sandwich ELISA Kit from Cell Signaling Technology is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected levels of tyrosine-phosphorylated Insulin Receptor β protein. An Insulin Receptor β Rabbit mAb has been coated on the microwells. After incubation with cell lysates, Insulin Receptor β protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine Mouse mAb is added to detect captured tyrosine-phosphorylated Insulin Receptor β protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate TMB is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of Insulin Receptor β protein phosphorylated on tyrosine.

Background

Insulin receptor (INSR) is a membrane receptor tyrosine kinase. The receptor molecule consists of a disulfide linked heterodimer. The α subunit is a 135 kDa extracellular fragment, and the β subunit is a 95 kDa fragment containing an extracellular domain, a single transmembrane domain, and an intracellular tyrosine kinase domain (1). Insulin ligand binding to this receptor results in receptor autophosphorylation and tyrosine kinase activation. INSR catalyzes the tyrosine phosphorylation of molecules such as IRS, Gab1, Shc, and Cbl, which further activate the downstream MAPK, PI3K, and TC10 pathways. This eventually leads to increases in glucose uptake and metabolism as well as cell growth (2,3). INSR has peptide substrate specificity similar to other receptor tyrosine kinase members, preferring acidic residues at the -1 to -4 positions and large hydrophobic amino acids at positions +1 and +3 (4).

1. Yip, C.C. and Ottensmeyer, P. (2003) J. Biol. Chem. 278, 27329-27332.
2. Saltiel, A.R. and Pessin, J.E. (2002) Trends Cell Biol. 12, 65-71.
3. Zick, Y. (2001) Trends Cell Biol. 11, 437-441.
4. Songyang, Z. and Cantley, L.C. (1995) TIBS 20, 470-475.

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).