PathScan? Total IκBα Sandwich ELISA Kit

• 产品编号：CST-7360C      品牌：CST       原厂货号：7360C
• 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® Total IkappaB-alpha Sandwich ELISA Kit能检测内源性总 IκBα蛋白的水平。使用PathScan® Phospho-IκBα (Ser32) Sandwich ELISA Kit #7355，能检测 TNF-α处理的HeLa细胞中丝氨酸（32位点）一个重要的磷酸化诱导——phospho-IκBα。然而，用Sandwich ELISA Kit #7360检测的总 IκBα蛋白水平（磷酸化与非磷酸化的）却没有发生改变（图1）。该试剂盒经公司内部实验证明能检测已表明物种的蛋白，但也许能检测其他物种中的类似蛋白。CST的 PathScan® Total IκBα Sandwich ELISA Kit是一种固相三明治酶联免疫吸附实验 (ELISA) ，它能测定内源性 IκBα蛋白水平。IκBα Mouse mAb 7361*抗体能被覆盖在微量培养板上。用细胞裂解液孵化后，磷酸化和非磷酸化的IκBα蛋白就能被原来覆盖的抗体捕获。接着反复清洗后加入足量的IκBα Antibody 7362* 以检测捕获的 IκBα蛋白。然后用HRP-linked anti-rabbit antibody #7074*识别已绑定的检测性抗体。加入HRP的底物——TMB用于显色。显色剂颜色的深浅与 总IκBα 的含量成比例。这个试剂盒中的*抗体是特异定制的。NF-κB/Rel 转录因子以活化状态出现于胞质中，并与抑制因子 IκB蛋白复合(1-3)。活化的发生是通过 IκBα蛋白丝氨酸（32、36位点）的磷酸化，接着蛋白酶调节的降解导致活化的NF-κB蛋白释放，并转移到细胞核内(3-7)。IκBα 蛋白的磷酸化和产生的Rel依赖性转录被细胞外信号的高度多元化群体激活，这些群体包括炎性细胞因子，生长因子和趋化因子。目前已经鉴定出在这些活化位点磷酸化 IκB蛋白的激酶 (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.
9. Chen, J. et al. (2008) J Cell Mol Med , .

原厂资料：

Specificity / Sensitivity

CST's PathScan® Total IkappaB-alpha Sandwich ELISA Kit detects endogenous levels of total IκBα protein. Using PathScan® Phospho- 
 IκBα (Ser32) 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 this Sandwich ELISA Kit #7360, remains 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® Total IκBα Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total IκBα 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, an IκBα Antibody 7362* is added to detect the captured IκBα 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 total IκBα 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.
9. Chen, J. et al. (2008) J Cell Mol Med , .

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)