| 货号 | 9105T |
| 描述 | The Phospho-Bad Antibody Sampler Kit provides an economical means to investigate the role of Bad protein in apoptosis. The kit contains primary and secondary antibodies to perform four Western blots with each antibody. |
| 目标/特异性 | Bad (D24A9) Rabbit mAb detects endogenous levels of total Bad protein. Phospho-Bad (Ser112) (40A9) Rabbit mAb detects endogenous levels of Bad only when phosphorylated at Ser112. The Ser112 nomenclature is based upon the mouse sequence. The analogous phosphorylation sites are Ser75 in human and Ser113 in rat. Phospho-Bad (Ser136) (185D10) Rabbit mAb detects immunoprecipitated or transfected levels of Bad protein only when phosphorylated at Ser136. The Ser136 nomenclature is based upon the mouse sequence. The analogous phosphorylation sites are Ser99 in human and Ser137 in rat. Phospho-Bad (Ser155) Antibody detects transfected levels of Bad only when phosphorylated at Ser155. The Ser155 nomenclature is based upon the mouse sequence. The analogous phosphorylation sites are Ser118 in human and Ser156 in rat. |
| 供应商 | CST |
| 背景 | 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). |
| 存放说明 | -20C |
| 参考文献 | Yang, E. et al. (1995) Cell 80, 285-291. Zha, J. et al. (1996) Cell 87, 619-628. Datta, S.R. et al. (1997) Cell 91, 231-241. Peso, L. et al. (1997) Science 278, 687-689. Bonni, A. et al. (1999) Science 286, 1358-1362. Tan, Y. et al. (1999) J. Biol. Chem. 274, 34859-34867. Harada, H. et al. (1999) Mol. Cell 3, 413-422. Tan, Y. et al. (2000) J. Biol. Chem. 275, 25865-25869. Lizcano, J. et al. (2000) Biochem. J. 349, 547-557. Datta, S. et al. (2000) Mol. Cell 6, 41-51. |
Western blot analysis of extracts from various cell lines using Bad (D24A9) Rabbit mAb #9239. Western blot免疫印记。用Bad (D24A9) Rabbit mAb #9239分析各种细胞系的细胞提取液。 | |
Western blot analysis of cell extracts from 293 cells transfected with Wild-type Bad, Bad (S112A/S136A), Bad (S155A) and treated with forskolin (30 µM), using Phospho-Bad (Ser155) Antibody #9297 (upper) and Bad Antibody #9292 (lower). Western blot免疫印记分析分别转染了野生型Bad、Bad (S112A/S136A)和 Bad (S155A)并经forskolin (30 µM)处理的293细胞的细胞提取液，所用抗体为 Phospho-Bad (Ser155) Antibody #9297 (上图) 和Bad Antibody #9292 (下图)。 | |
Western blot analysis of extracts from control 293, GST-Bad transfected 293 and COS cells, untreated or TPA-treated, using Phospho-Bad (Ser136) (185D10) Rabbit mAb #5286. Western blot 免疫印记。用Phospho-Bad (Ser136) (185D10) Rabbit mAb 兔单抗#5286分析对照293细胞, 转染了GST-Bad 的293细胞和COS细胞的细胞提取液，并未经过处理和经过TPA处理。 | |
Western blot analysis of extracts from COS cells, untreated or TPA-treated, using Phospho-Bad (Ser112) (40A9) Rabbit mAb #5284 (upper) or Bad Antibody #9292 (lower). Western blot免疫印记。用Phospho-Bad (Ser112) (40A9) Rabbit mAb 兔单抗#5284 (上图) 或者Bad Antibody 多抗#9292 (下图)分析未经处理的或经TPA处理的COS细胞的细胞提取液。 | |
Western blot analysis of lysates containing Bad Control Proteins #9293, comprised of nonphosphorylated and phosphorylated Bad peptides, using Phospho-Bad (Ser136) (D25H8) Rabbit mAb. | |
Western blot analysis of extracts from COS-7 cells, treated with Human Epidermal Growth Factor #8916 (100 ng/ml, 30 min) in the presence or absence of calf intestinal phosphatase (CIP) plus lambda-phosphatase, using Phospho-Bad (Ser136) (D25H8) Rabbit mAb (upper) or Bad (D24A9) Rabbit mAb #9239 (lower). | |
Western blot analysis of extracts from ACHN cells, untreated or treated with Human Epidermal Growth Factor #8916 (100 ng/ml, 30 min), and C2C12 cells, untreated or insulin-treated (100 nM, 30 min), using Phospho-Bad (Ser136) (D25H8) Rabbit mAb (upper) or Bad (D24A9) Rabbit mAb #9239 (lower). | |
Western blot analysis of extracts from various cell lines using Bad (D24A9) Rabbit mAb. | |
Western blot analysis of HeLa cells, mock transfected or transfected with pCMV-Tag4A-mBad/GrpE, using Bad (D24A9) Rabbit mAb #9239. 使用 Bad (D24A9) Rabbit mAb 兔单抗#9239 对转染了对照或pCMV-Tag4A-mBad/GrpE的HeLa细胞的细胞提取液进行 Western blot 免疫印记分析。 | |
Western blot analysis of HeLa cells, mock transfected or transfected with pCMV-Tag4A-mBad/GrpE, untreated or TPA- and/or λ-phosphatase-treated, using phospho- and total Bad antibodies #5286, #9296, #9297, and #9268. Note that overexpressed fusion protein runs as a 48 to 52 kDa doublet that is likely formed as a result of alternative start sites. Western blot 免疫印记检测分别转染了对照或pCMV-Tag4A-mBad/GrpE的HeLa细胞的细胞提取液，分为未处理组和TPA-和/或λ-磷酸酶处理组，分别用phospho-Bad 和 total Bad antibodies #5286、#9296、#9297和#9268 进行分析。需要注意的是，过表达的融合蛋白电泳时出现在48 到 52 kDa之间 的双条带，很有可能是可变起始位点引起的。 | |
Flow cytometric analysis of COS cells, untreated (blue) or TPA/Calyculin A treated (green), using Phospho-Bad (Ser112) (40A9) Rabbit mAb. | |
Western blot analysis of extracts from COS cells, untreated or TPA-treated, using Phospho-Bad (Ser112) (40A9) Rabbit mAb (upper) or Bad Antibody #9292 (lower). | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-Bad (Ser 112) (40A9) Rabbit mAb. | |
Immunohistochemical analysis of paraffin embedded COS cells untreated (left) or TPA-treated (right), showing induced cytoplasmic staining using Phospho-Bad (Ser112) (40A9) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded Non-Hodgkins lymphoma, using Phospho-Bad (Ser112) (40A9) Rabbit mAb. |
