| 货号 | 9271S |
| 反应种属 | Human/Mouse/Rat/Hamster/D.melanogaster/Bovine/Dog/Pig |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/F |
| 目标/特异性 | Phospho-Akt (Ser473) Antibody detects endogenous levels of Akt1 only when phosphorylated at Ser473. This antibody also recognizes Akt2 and Akt3 when phosphorylated at the corresponding residues. It does not recognize Akt phosphorylated at other sites, nor does it recognize phosphorylated forms of related kinases such as PKC or p70 S6 kinase. |
| 使用方法 | WB(1:1000) IP (1:100) F (1:50) IF-IC (1:25) |
| 供应商 | CST |
| 背景 | Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip (15) and p21 Waf1/CIP1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19). |
| 存放说明 | -20C |
| 计算分子量 | 60 |
Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with PDGF, wortmannin, LY294002, rapamycin or PD98059, using Phospho-Akt (Ser473) Antibody. Western blot 分析NIH/3T3 细胞提取物, 非处理组和PDGF, wortmannin, LY294002, rapamycin 或PD98059处理组，所用抗体为 Phospho-Akt (Ser473) Antibody。 | |
Western blot analysis of immunoprecipitated Akt from 293 cells transiently transfected with HA-tagged Akt (WT), HA-tagged K179A mutant Akt and HA-tagged K179A/S473A mutant Akt, using Phospho-Akt (Ser473) Antibody (upper), Akt antibody (middle) or HA antibody (lower). Phospho-Akt (Ser473) Antibody does not recognize Akt with an alanine substituion at Ser473. (Polakiewicz, R.D. et al. [1998] J. Biol. Chem. 273, 23534-23541.) Western blot 分析瞬时转染HA-tagged Akt (WT), HA-tagged K179A突变Akt 和 HA-tagged K179A/S473A 突变 Akt的293 细胞免疫沉淀的Akt，所用抗体为Phospho-Akt (Ser473) Antibody (上), Akt antibody (中) or HA antibody （下)。 Phospho-Akt (Ser473) Antibody 不能识别丙氨酸代换的Akt。(Polakiewicz, R.D. et al. [1998] J. Biol. Chem. 273, 23534-23541.) | |
Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with PDGF for the indicated times, using Phospho-Akt (Ser473) Antibody (upper) or Akt Antibody #9272 (lower). Western blot分析NIH/3T3细胞的提取物，分为未处理组和PDGF处理组，如图中的时间所示，使用的抗体是Phospho-Akt (Ser473) Antibody (上) 或者 Akt Antibody #9272 (下)。 | |
Flow cytometric analysis of LNCaP cells, untreated (green) or LY294002-treated (blue), using Phospho-Akt (Ser473) Antibody compared to a nonspecific negative control antibody (red). 流式细胞术分析未处理（绿色）或者LY294002处理（蓝色）的LNCaP细胞，使用的抗体是Phospho-Akt (Ser473) Antibody，并和非特异性阴性对照抗体比较（红色）。 | |
Confocal immunofluorescent images of C2C12 cells serum starved and treated with or without insulin as indicated and labeled with Phospho-Akt (Ser473) Antibody (red). Actin filaments have been labeled with fluorescein phalloidin. 免疫荧光共聚焦分析C2C12细胞，如图分别为血清饥饿，胰岛素未处理或者处理组，并应用Phospho-Akt (Ser473) 抗体标记（红色）。肌动蛋白纤维由荧光素-鬼笔环肽（fluorescein phalloidin）所标记。 |
