PathScan? Total IRS-2 Sandwich ELISA Kit

• 产品编号：CST-7884C      品牌：CST       原厂货号：7884C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

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描述：

Sandwich ELISA Kit #7884进行检测，在转染的CHO细胞中，IRS-2表达水平明显上调。Western检测显示，IRS-2总蛋白(磷酸化与非磷酸化)水平在胰岛素处理后没有改变。该试剂盒同样可以用于检测3T3-L1脂肪细胞中内源性的IRS-2水平(数据未列出)。内部测试证明，该试剂盒除了可以检测相应物种的相关蛋白外，也可与其它物种的同源蛋白交互反应。PathScan® Total IRS-2 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IRS-2(也称作4PS)总蛋白。步骤如下：首先在 微孔中结合有固相的兔抗IRS-2抗体，然后将细胞裂解液于固相抗体中孵育，IRS-2(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，然后加入生物素化的兔源IRS-2检测抗体来检测被捕获的IRS-2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。IRS-2的浓度与显色的强度成正比，因此可以借此对IRS-2定量。胰岛素受体底物2(IRS-2)是胰岛素受体激酶的主要底物之一(1)。在脊椎动物中， IRS-2主要作为一个支架蛋白，用于协调Insulin/IGF信号级联反应中的各个分支通路(2)。IRS-2对于正常的营养稳态是必需的，因为它不仅倡导外周组织中的胰岛素功能，还调节着IGF-1对于B细胞生长的作用。IRS-2缺失的小鼠不能维持足够量的胰岛素代偿性分泌，并且容易患1型糖尿病(3)。

1. Sun, X.J. et al. (1991) Nature 352, 73-77.
2. White, M.F. (2002) Am. J. Physiol. Endocrinol. Metab. 283, E413-E422.
3. Withers, D.J. et al. (1998) Nature 391, 900-904.

原厂资料：

Specificity / Sensitivity

CST''s PathScan® Total IRS-2 Sandwich ELISA Kit #7884 detects endogenous levels of IRS-2. As shown in Figure 1, using the PathScan® Total IRS-2 Sandwich ELISA Kit #7884, a high level of IRS-2 is detected in transfected CHO cells. The total levels of IRS-2 (phospho and nonphospho) remain unchanged after insulin treatment as shown by western analysis. This kit also detects endogenous levels of IRS-2 expressed by 3T3-L1 adipocytes (data not shown). 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® Total IRS-2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total IRS-2 (also called 4PS). An IRS-2 rabbit antibody has been coated onto the microwells. After incubation with cell lysates, IRS-2 (both phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a biotinylated IRS-2 rabbit antibody is added to detect the captured IRS-2. HRP-linked streptavidin 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 IRS-2.

Background

Insulin Receptor Substrate 2 (IRS-2) is one of the major substrates of the insulin receptor kinase (1). In vertebrates, IRS-2 functions as a scaffolding protein to coordinate separate branches of the Insulin/IGF-signaling cascades (2). IRS-2 is essential for normal nutrient homeostasis because it mediates both peripheral insulin action and the effect of IGF-1 on B-cell growth. Mice lacking IRS-2 fail to maintain sufficient compensatory insulin secretion and develop diabetes as young adults (3).

1. Sun, X.J. et al. (1991) Nature 352, 73-77.
2. White, M.F. (2002) Am. J. Physiol. Endocrinol. Metab. 283, E413-E422.
3. Withers, D.J. et al. (1998) Nature 391, 900-904.

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).