PathScan? Multiplex Western Cocktail I: Phospho-p90RSK, Phospho-Akt, Phospho-p44/42 MAPK (Erk1/2) an

• 产品编号：CST-5301S      品牌：CST       原厂货号：5301S
• 产品分类：抗体 > 一抗 > 磷酸化抗体
• 应用分类：

描述：

该cocktail中的每个磷酸化抗体都能识别内源性特异靶点磷酸化的蛋白水平。Rab11 Antibody都能检测总它的靶蛋白水平，并且不依赖于磷酸化，同时该抗体还能用于蛋白负载对照。该多克隆抗体通过使用合成肽段免疫动物而获得。该抗体经蛋白A和肽亲和层析纯化。PathScan® Multiplex Western Cocktail I为分析一张膜上各种蛋白的活化状态提供了独特的方法，不用去除免疫印迹膜上的封闭液和抗体，然后再次重新标记。该方法为使用者节约了时间，且调高了准确度，减少了试剂的浪费。该系统让使用者能即时检测phospho-p90RSK, phospho-Akt, phospho-p44/42 MAPK (Erk1/2) 和phospho-S6核糖体蛋白。Cocktail也包括用于控制蛋白负载的Rab11 Antibody。Akt也被称为PKB或Rac，它在控制生存和凋亡平衡中起着重要作用(1-3)。该蛋白激酶被胰岛素、各种生长、存活因子活化，并且作用于涉及PI3激酶的渥曼青霉素敏感性通路(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。 丝氨酸（473位点）羧端的磷酸化也能激活Akt。p44 and p42 MAP kinases (Erk1 and Erk2)在调节细胞生长分化中起了重要作用5-8)。许多细胞外信号分子都能激活MAP kinases ，包括生长、神经营养因子，细胞因子，激素和神经传导物质。MAP kinases活化的发生通过单个上游MAP激酶的激酶(MEK)对 T*EY*序列中苏氨酸和酪氨酸(人源MAP kinase [Erk1] 蛋白202、204位点和大鼠Erk2蛋白183、185位点)的磷酸化(9,10)而实现。p44/42 MAPK的其中一个下游靶点是p90RSK。为了有效地促进可持续的方式中细胞的生长和分裂，生长因子和有丝分裂原必须上调转移（11,12)。生长因子和有丝分裂原诱导p70 S6 kinase的活化，该激酶又能磷酸化S6核糖体蛋白。S6的磷酸化与转位增加有关，特别是5+非转位区域内带帽结构的mRNAs(12)。

1. Franke, T.F. (1997) Cell 88, 435-437.
2. Burgering, B.T. and Coffer, P.J. (1995) Nature 376, 599-602.
3. Franke, T. F. et al. (1995) Cell 81, 727-736.
4. Alessi, D.R. et al. (1996) EMBO J. 15, 6541-6551.
5. Marshall, C.J. (1995) Cell 80, 179-185.
6. Hunter, T. (1995) Cell 80, 225-236.
7. Hill, C.S. and Treisman, R. (1995) Cell 80, 199-211.
8. Cowley, S. et al. (1994) Cell 77, 841-852.
9. Sturgill, T.W. et al. (1988) Nature 334, 715-718.
10. Payne, D. M. et al. (1991) EMBO J. 10, 885-892.
11. Dufner, A. and Thomas, G. (1999) Exp. Cell. Res. 253, 100-109.
12. Peterson, R.T. and Schreiber, S.L. (1998) Curr. Biol. 8, R248-R250.

原厂资料：

Specificity / Sensitivity

Each phospho-antibody in this cocktail recognizes endogenous levels of only the phosphorylated form of its specific target. The Rab11 antibody detects endogenous levels of its target protein independent of phosphorylation, and is provided to control for protein loading.

Source / Purification

Antibodies are produced by immunizing animals with synthetic peptides. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Description

The PathScan® Multiplex Western Cocktail I offers a unique method to assay the activation of multiple pathways on one membrane without stripping and reprobing. This method saves the user valuable time, while increasing accuracy and minimizing reagent waste. The system allows the user to simultaneously detect levels of phospho-p90RSK, phospho-Akt, phospho-p44/42 MAPK (Erk1/2) and phospho-S6 ribosomal protein. The cocktail also includes the Rab11 Antibody to control for protein loading.

Background

Akt, also referred to as PKB or Rac, plays a critical role in controlling the balance between survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors, and functions 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 also by phosphorylation within the carboxy-terminus at Ser473. 
 Both p44 and p42 MAP kinases (Erk1 and Erk2) play a critical role in the regulation of cell growth and differentiation (5-8). MAP kinases are activated by a wide variety of extracellular signals including growth and neurotrophic factors, cytokines, hormones and neurotransmitters. Activation of MAP kinases occurs through phosphorylation of threonine and tyrosine (202 and 204 of human MAP kinase [Erk1] or 183 and 185 of rat Erk2) at the sequence T*EY* by a single upstream MAP kinase kinase (MEK) (9,10). One of the downstream targets of p44/42 MAPK is p90RSK. 
 To effectively promote growth and cell division in a sustained manner, growth factors and mitogens must upregulate translation (11,12). Growth factors and mitogens induce the activation of p70 S6 kinase, which in turn phosphorylates the S6 ribosomal protein. Phosphorylation of S6 correlates with an increase in translation, particularly of mRNAs with an oligopyrimidine tract in their 5+ untranslated regions (12).

1. Franke, T.F. (1997) Cell 88, 435-437.
2. Burgering, B.T. and Coffer, P.J. (1995) Nature 376, 599-602.
3. Franke, T. F. et al. (1995) Cell 81, 727-736.
4. Alessi, D.R. et al. (1996) EMBO J. 15, 6541-6551.
5. Marshall, C.J. (1995) Cell 80, 179-185.
6. Hunter, T. (1995) Cell 80, 225-236.
7. Hill, C.S. and Treisman, R. (1995) Cell 80, 199-211.
8. Cowley, S. et al. (1994) Cell 77, 841-852.
9. Sturgill, T.W. et al. (1988) Nature 334, 715-718.
10. Payne, D. M. et al. (1991) EMBO J. 10, 885-892.
11. Dufner, A. and Thomas, G. (1999) Exp. Cell. Res. 253, 100-109.
12. Peterson, R.T. and Schreiber, S.L. (1998) Curr. Biol. 8, R248-R250.

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 the antibody.