PhosphoPlus? Akt (Ser473) In-Cell Duet (ICW Compatible)

• 产品编号：CST-7255S      品牌：CST       原厂货号：7255S
• 产品分类：抗体 > 一抗 > 抗体对
• 应用分类：

其它组分：

描述：

Phospho-Akt (Ser473) antibody可以检测丝氨酸473位点磷酸化的内源性的Akt蛋白。总Akt 检测内源性的Akt总蛋白水平，此抗体不与其它相关蛋白发生交叉反应。单克隆抗体采用合成的与人Akt丝氨酸473位点周围或羧基末端序列相对应的磷酸肽段免疫动物生产的。CST公司的PhosphoPlus® Akt (Ser473) In-Cell Duet为研究蛋白的活化状态提供了一种简便的方法，它采用具有近红外的探测能力的多孔板扫描仪器进行检测，如LI-COR® Biosciences Odyssey® 红外成像系统。本试剂盒内含经过高标准的遴选的总蛋白及已优化的处于活化状态的的抗体混合物。总蛋白和磷酸化的抗体可同时在一个孔中检测出来，使得磷酸化蛋白的水平可与总蛋白进行归一化。近红外检测混合也包含在其中。 Akt,又被称为PKB 或 Rac，在细胞的生长和凋亡中起到关键作用(1-3)。被胰岛素（insulin）或各种生长及存活因子激活后，这个激酶在包括PI3K在内的对wortmannin敏感的通路中起作用(2,3)。磷脂结合可以调控Akt的激活，其苏氨酸308位点被PDK1环磷酸化(4)，并且在该蛋白碳链末端丝氨酸（473）位点磷酸化。以前研究认为PDK2在丝氨酸473位点磷酸化Akt，实际为哺乳类动物雷帕霉素靶蛋白（mTOR，mammalian target of rapamycin），包含rictor 和Sin1 的雷帕霉素非敏感复合体作用该位点。Akt促进细胞的生长通过抑制细胞的凋亡 ，例如Akt可以抑制下游靶蛋白Bad (7), forkhead 转录因子 (8), c-Raf (9), and caspase-9。PTEN 磷酸酶是抑制PI3 激酶/Akt信号通路的主要蛋白(10)。 LY294002是特异的 PI3 激酶的抑制剂(11)。Akt 的另外一个主要功能是通过磷酸化和抑制GSK-3α 和 β调控糖原的合成(12,13)。Akt 也可以调控insulin诱导的葡萄糖转运 (12)。除此之外，Akt还可以调控细胞周期，通过抑制GSK-3β，从而调控其下游的cyclin D1 的磷酸化和降解 (14)，或者负向调控cyclin依赖的激酶抑制因子 p27 Kip (15) and p21 Waf1/CIP1 (16)。Akt 也可以通过直接磷酸化mTOR（其在包含raptor的雷帕霉素敏感复合体内）在调控细胞的生长其起重要作用(17)。更重要的是，Akt磷酸化并抑制tuberin（TSC2）， 而TSC2是mTOR-raptor 复合体中的mTOR的抑制因子(18,19)。抑制mTOR会导致p70 S6激酶失活并激活翻译的抑制因子4E-BP1（真核起始因子4E结合蛋白1），从而终止蛋白合成（18，19）。

原厂资料：

Specificity / Sensitivity

Phospho-Akt (Ser473) antibody detects endogenous levels of Akt only when phosphorylated at Ser473. Total Akt detects endogenous levels of total Akt protein. This antibody does not cross-react with other related proteins.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser473 of human Akt or at the carboxy terminal sequence of human Akt.

Description

PhosphoPlus® Akt (Ser473) In-Cell Duet from Cell Signaling Technology (CST) provides an easy method to assess protein activation status using a multi-well plate scanner with near infrared detection capabilities, such as the LI-COR® Biosciences Odyssey® Infrared Imaging System. This kit contains a pre-optimized activation-state and total protein antibody cocktail, selected based on superior performance. Phosphorylated and total protein are detected simultaneously in the same well, allowing levels of phosphorylated protein to be normalized to total protein. A near infrared detection cocktail is also included. 

Background

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). Inhibition of mTOR stops the protein synthesis machinery by inactivating p70 S6 kinase and activating the eukaryotic initiation factor 4E binding protein 1 (4E-BP1), an inhibitor of translation (18,19). 

注意事项：

Storage: Supplied in 10 mM sodium HEPES
(pH 7.5), 150 mM, NaCl, 100 μg/ml BSA and 50% glycerol.
Store at –20°C. Do not aliquot either cocktail.