PathScan? Phospho-Akt1 (Ser473) Sandwich ELISA Antibody Pair

• 产品编号：CST-7143S      品牌：CST       原厂货号：7143S
• 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性，请参考相应的PathScan® Sandwich ELISA试剂盒。注意：经根据内部测试结果，此抗体可检测指定物种的蛋白，但也可能检测其他物种同源性蛋白。PathScan ® Phospho-Akt1 (Ser473) Sandwich ELISA Antibody Pair是CST提供的对于PathScan ® Phospho-Akt1 (Ser473) Sandwich ELISA Kit #7160的另外一种经济的选择。其内有捕获和检测抗体(100X )以及ＨＲＰ标记(1000X )的二抗。足够量的试剂可以用来做４个９６孔板的Elisa。兔源Phospho-Akt (Ser473)抗体在96微孔板中PBS包被过夜。封闭后，加入细胞的裂解液共孵育， 然后加入鼠源AKT1抗体和HRP标记抗鼠IgG的抗体。加入ＨＲＰ底物（ＴＭＢ）用以显色。所产生颜色的吸光度值与phospho-Akt1 (Ser473)蛋白的含量呈正相关。*试剂盒中的抗体是为此试剂盒特别定制的。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)。该蛋白激酶可以被胰岛素和多种生长和存活因子激活，在涉及PI3K激酶的wortmannin敏感信号通路中发挥作用（2,3）。Akt可由磷脂结合激活，该过程通过活化环中的Thr308位点以及羧基端Ser473位点的磷酸化完成，其中Thr308的磷酸化由PDK1完成(4)。 之前推测PDK2在Ser473位点磷酸化Akt，后被证实为哺乳动物rapamycin靶蛋白mTOR 的作用， 它存在在一个含有rictor和Sin1 的rapamycin非敏感复合体中。 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是PI3K/Akt信号通路的主要负调控因子(10)。 LY294002是特异性的 PI3K 激酶抑制剂(11)。Akt 的另外一个主要功能是通过磷酸化并进而抑制GSK-3α 和 β来调控糖原的合成(12,13)。 Akt 也可以调控胰岛素诱导的葡萄糖转运 (12)。除此之外，Akt还可以调控细胞周期，这个功能通过抑制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) 和 p21 Waf1/CIP1 (16)来实现。 Akt 也可以通过直接磷酸化含有raptor的rapamycin敏感复合体中的mTOR来调控细胞的生长(17)。 更重要的是， Akt磷酸化并失活TSC2， 而TCS2是mTOR-raptor复合物中mTOR的抑制因子(18,19)。

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-Akt1 (Ser473) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-Akt1 (Ser473) Sandwich ELISA Kit #7160. 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 Phospho-Akt (Ser473) Rabbit Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by Akt1 Mouse Detection Antibody and HRP-conjugated Anti-Mouse IgG. HRP substrate (TMB) is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-Akt1 (Ser473) protein. 
 *Antibodies in this kit are custom formulations specific to the kit.

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 (18,19).

注意事项：

Storage: Phospho-Akt (Ser 473) Rabbit Capture and Akt1
Mouse Detection Antibodies are stored at 4°C.
Anti-Mouse IgG-HRP Linked Antibody is stored at -20°C