PathScan? Phospho-IκBα (Ser32) Sandwich ELISA Kit

• 产品编号：CST-7355C      品牌：CST       原厂货号：7355C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

其它组分：

描述：

CST的PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit 检测内源水平在Ser32位点磷酸化的IκBα蛋白。如图1所示,用Sandwich ELISA Kit #7355能够检测到经TNF-α诱导的HeLa细胞中在Ser32位点磷酸化的IκBα。然而，用Total IκBα Sandwich ELISA Kit #7360检测总IκBα (磷酸化和非磷酸化)的蛋白水平并未发生变化。此试剂盒能够检测已经经内部严格测试的物种的蛋白，但是也不排除检测到其它物种中同源性的蛋白。CST的PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit 是固相夹心酶联免疫吸收剂检测试剂盒(ELISA)，检测内源水平在Ser32位点磷酸化的IκBα蛋白。IκBα Mouse mAb #7361*包裹到微孔的外表面。然后与细胞裂解液孵育，磷酸化和非磷酸化的IκBα蛋白能够被抗体捕获。过严格的冲洗后，Phospho-IκBα (Ser32) Antibody #2859*加入用来检测捕捉到的phospho-IκBα (Ser32)蛋白。HRP连接抗兔抗体#7074*用于识别检测抗体的结合。HRP的底物TMB加入进行显色反应。对反应的颜色，吸光度的数值按比例换算成phospho-IκBα (Ser32)蛋白的量。* 试剂盒中的抗体是经过客户验证的特异性抗体。NF-κB/Rel转录调控因子在细胞质中与IκB抑制蛋白结合以非活性形式存在(1-3)。磷酸化IκBα 的Ser32 和 Ser36位点后，蛋白水解酶体介导的IκB降解使得 NF-κB释放激活并入核(3-7)。IκBα的磷酸化导致了Rel依赖的转录能够被很多胞外信号激活，包括炎症因子，生长因子和趋化因子 。能够磷酸化IκB这些活性位点的激酶已经得到了鉴定(8)。

1. Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6.
2. Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70.
3. Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8.
4. Brown, K. et al. (1995) Science 267, 1485-8.
5. Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18.
6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
7. Chen, Z.J. et al. (1996) Cell 84, 853-62.
8. Karin, M. and Ben-Neriah, Y. (2000) Annu Rev Immunol 18, 621-63.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit detects endogenous levels of Phospho-IκBα (Ser32) protein. As shown in Figure 1, using this Sandwich ELISA Kit #7355, a significant induction of Phospho-IκBα (Ser32) in HeLa cells treated with TNF-α can be detected. However, the level of total IκBα (phospho- and nonphospho-), detected by the Total IκBα Sandwich ELISA Kit #7360, remains unchanged. 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-IκBα (Ser32) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Phospho-IκBα (Ser32) protein. An IκBα Mouse mAb #7361* has been coated onto the microwells. After incubation with cell lysates, both nonphospho- and phospho-IκBα proteins are captured by the coated antibody. Following extensive washing, Phospho-IκBα (Ser32) Antibody #2859* is added to detect the captured phospho-IκBα (Ser32) protein. HRP-linked anti-rabbit antibody #7074* 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-IκBα (Ser32) protein. * Antibodies in kit are custom formulations specific to kit.

Background

The NF-κB/Rel transcription factors are present in the cytosol in an inactive state complexed with the inhibitory IκB proteins (1-3). Activation occurs via phosphorylation of IκBα at Ser32 and Ser36 followed by proteasome-mediated degradation that results in the release and nuclear translocation of active NF-κB (3-7). IκBα phosphorylation and resulting Rel-dependent transcription are activated by a highly diverse group of extracellular signals including inflammatory cytokines, growth factors, and chemokines. Kinases that phosphorylate IκB at these activating sites have been identified (8).

1. Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6.
2. Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70.
3. Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8.
4. Brown, K. et al. (1995) Science 267, 1485-8.
5. Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18.
6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
7. Chen, Z.J. et al. (1996) Cell 84, 853-62.
8. Karin, M. and Ben-Neriah, Y. (2000) Annu Rev Immunol 18, 621-63.

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).