PathScan? Phospho-Bad (Ser112) Sandwich ELISA Antibody Pair

• 产品编号：CST-7842S      品牌：Cell Signaling Technology       原厂货号：7842S
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测抗体对
• 应用分类：

其它组分：

描述：

抗体对的特异性和灵敏度请参考PathScan® Sandwich ELISA Kit。注意：经过严格的内部实验验证，此抗体对能够检测指定物种的蛋白。但是可能也能检测其它物种的同源蛋白。该CST生产的PathScan® Phospho-Bad (Ser112) Sandwich ELISA Antibody Pair 是本公司PathScan® Phospho-Bad (Ser112) Sandwich ELISA Kit #7182的经济型的替代产品。捕获抗体和检测抗体(100X 存储液)和HRP偶联的二抗(1000X 存储液)都有提供，并且其中的试剂足够用于4 x 96孔ELISAs所需的量。Bad捕获抗体包被在96孔板中，加上PBS溶液孵育过夜。封闭以后，加入细胞裂解液，随后加入Phospho-Bad (Ser112) 检测抗体和抗鼠IgG、HRP偶联抗体，最后加入HRP底物，TMB显色。颜色反应的吸光度数值按照比例换算成Phospho-Bad (Ser112)蛋白的总量。 *试剂盒中的抗体是为该试剂盒特制的。Bad是Bcl-2促凋亡家族中的成员，它通过转移 Bax 与 Bcl-2 和 Bcl-xL 结合而促进细胞死亡(1,2)。存活因子如IL-3, 能够通过激活细胞内信号通路，导致Bad在Ser112和Ser136位点发生磷酸化，进而抑制凋亡活性(2)。在这些位点的磷酸化促进Bad与 14-3-3 蛋白的结合，从而阻止了Bad与Bcl-2和Bcl-xl的互作(2)。Akt在Ser136位点磷酸化Bad，而促进细胞的存活(3,4)。Bad在体内和体外都能够被p90RSK(5,6)和线粒体锚定PKA磷酸化Ser112位点（7）。PKA对BH3结构域Ser155位点的磷酸化，在阻止Bad和Bcl-xL的二聚化过程中发挥了重要的作用(8-10)。

1. Yang, E. et al. (1995) Cell 80, 285-291.
2. Zha, J. et al. (1996) Cell 87, 619-628.
3. Datta, S.R. et al. (1997) Cell 91, 231-241.
4. Peso, L. et al. (1997) Science 278, 687-689.
5. Bonni, A. et al. (1999) Science 286, 1358-1362.
6. Tan, Y. et al. (1999) J. Biol. Chem. 274, 34859-34867.
7. Harada, H. et al. (1999) Mol. Cell 3, 413-422.
8. Tan, Y. et al. (2000) J. Biol. Chem. 275, 25865-25869.
9. Lizcano, J. et al. (2000) Biochem. J. 349, 547-557.
10. Datta, S. et al. (2000) Mol. Cell 6, 41-51.

原厂资料：

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair 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-Bad (Ser112) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-Bad (Ser112) Sandwich ELISA Kit #7182. Capture and Detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Bad Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a Phospho-Bad (Ser112) Detection Antibody and an anti-Mouse IgG, HRP conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of Phospho-Bad (Ser112) protein. 
 *Antibodies in this kit are custom formulations specific to the kit.

Background

Bad is a proapoptotic member of the Bcl-2 family that promotes cell death by displacing Bax from binding to Bcl-2 and Bcl-xL (1,2). Survival factors, such as IL-3, inhibit the apoptotic activity of Bad by activating intracellular signaling pathways that result in the phosphorylation of Bad at Ser112 and Ser136 (2). Phosphorylation at these sites promotes binding of Bad to 14-3-3 proteins to prevent an association between Bad with Bcl-2 and Bcl-xl (2). Akt phosphorylates Bad at Ser136 to promote cell survival (3,4). Bad is phosphorylated at Ser112 both in vivo and in vitro by p90RSK (5,6) and mitochondria-anchored PKA (7). Phosphorylation of Ser155 in the BH3 domain by PKA plays a critical role in blocking the dimerization of Bad and Bcl-xL (8-10).

1. Yang, E. et al. (1995) Cell 80, 285-291.
2. Zha, J. et al. (1996) Cell 87, 619-628.
3. Datta, S.R. et al. (1997) Cell 91, 231-241.
4. Peso, L. et al. (1997) Science 278, 687-689.
5. Bonni, A. et al. (1999) Science 286, 1358-1362.
6. Tan, Y. et al. (1999) J. Biol. Chem. 274, 34859-34867.
7. Harada, H. et al. (1999) Mol. Cell 3, 413-422.
8. Tan, Y. et al. (2000) J. Biol. Chem. 275, 25865-25869.
9. Lizcano, J. et al. (2000) Biochem. J. 349, 547-557.
10. Datta, S. et al. (2000) Mol. Cell 6, 41-51.

注意事项：

Storage: Capture and Detection Antibodies are stored at 4ºC.
Anti-Mouse IgG, HRP-Linked Antibody is stored at -20ºC.