PathScan? Phospho-IKKβ (Ser177/181) Sandwich ELISA Kit

• 产品编号：CST-7080C      品牌：CST       原厂货号：7080C
• 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：

PathScan® Phospho-IKKβ (Ser177/181) Sandwich ELISA Kit检测内源水平在Ser177/181位点磷酸化的IKKβ蛋白，如图1所示。试剂盒的灵敏性如图2所示。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。PathScan® Phospho-IKKβ (Ser177/181) 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Ser177/181位点磷酸化的IKKβ蛋白。phospho-IKKβ (Ser177/181)兔抗包裹到微孔的外表面。然后与细胞裂解液孵育，phospho-IKKβ (Ser177/181)蛋白能够被抗体捕获。经过严格的冲洗后，IKKβ鼠检测抗体加入用来检测捕捉到的IKKβ蛋白。抗鼠IgG, HRP连接抗体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Ser177/181位点磷酸化的IKKβ蛋白的量。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)。大部分刺激因素采用通常信号通路即磷酸化诱导，蛋白水解酶体介导IκB的降解方式激活NF-κB (3-7)。此通路中最重要的步骤是激活高分子量的IκB激酶（IKK）复合体，此复合体催化的反应中有三个紧密相关的IKK亚基参与完成，IKKα and IKKβ 作为激酶的催化单元，IKKγ作为调控单元(8,9)。IKK的激活依赖于特定位点的磷酸化：IKKβ活性环中的 Ser177和Ser181位点以及IKKα中的serine 176 和 180 位点，这些位点的磷酸化引起蛋白构象的变化从而激活激酶（10-13）。

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. Zandi, E. et al. (1997) Cell 91, 243-52.
9. Karin, M. (1999) Oncogene 18, 6867-74.
10. DiDonato, J.A. et al. (1997) Nature 388, 548-54.
11. Mercurio, F. et al. (1997) Science 278, 860-6.
12. Johnson, L.N. et al. (1996) Cell 85, 149-58.
13. Delhase, M. et al. (1999) Science 284, 309-13.

原厂资料：

Specificity / Sensitivity

The PathScan® Phospho-IKKβ (Ser177/181) Sandwich ELISA Kit detects endogenous levels of IKKβ protein when phosphorylated at Ser177/181, 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-IKKβ (Ser177/181) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IKKβ when phosphorylated at Ser177/181. A phospho-IKKβ (Ser177/181) rabbit mAb has been coated onto the microwells. After incubation with cell lysates, phospho-IKKβ (Ser177/181) protein is captured by the coated antibody. Following extensive washing, an IKKβ mouse detection mAb is added to detect the captured IKKβ 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 the developed color is proportional to the quantity of IKKβ phosphorylated at Ser177/181.

Background

The NF-κB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IκB proteins (1-3). Most agents that activate NF-κB do so through a common pathway based on phosphorylation-induced, proteasome-mediated degradation of IκB (3-7). The key regulatory step in this pathway involves activation of a high molecular weight IκB kinase (IKK) complex, whose catalysis is generally carried out by three tightly associated IKK subunits. IKKα and IKKβ serve as the catalytic subunits of the kinase and IKKγ serves as the regulatory subunit (8,9). Activation of IKK depends upon phosphorylation; Ser177 and Ser181 in the activation loop of IKKβ (serine 176 and 180 in IKKα) are the specific sites whose phosphorylation causes conformational changes resulting in kinase activation (10-13).

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. Zandi, E. et al. (1997) Cell 91, 243-52.
9. Karin, M. (1999) Oncogene 18, 6867-74.
10. DiDonato, J.A. et al. (1997) Nature 388, 548-54.
11. Mercurio, F. et al. (1997) Science 278, 860-6.
12. Johnson, L.N. et al. (1996) Cell 85, 149-58.
13. Delhase, M. et al. (1999) Science 284, 309-13.

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).