PathScan? Multiplex Western Cocktail II: Phospho-p90RSK, Phospho-p53, Phospho-p38 MAPK and Phospho-S

• 产品编号：CST-5302S      品牌：CST       原厂货号：5302S
• 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。eIF4E Antibody都能检测它的总靶蛋白水平，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 IIPathscan® Multiplex Western Cocktail II为分析一张膜上各种蛋白的活性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-p53, phospho-p38 MAPK和phospho-S6核糖体蛋白。该试剂盒也包括用于控制蛋白负载的elF4E Antibody。90 kDa核糖体S6激酶(RSK1-3)是一类丝氨酸/苏氨酸激酶家族，广泛表达于许多生长因子、多肽激素和神经传导物质的应答反应之后(1)。p90RSK在体外能被Erk1、Erk2能激活，在体内能通过磷酸化而激活(2)。一些位点如丝氨酸（380、363位点）、苏氨酸（359位点），它的活化十分重要(3)。p53肿瘤抑制子蛋白在细胞应答DNA损伤和其他基因组紊乱中起着主要作用。p53的活化导致细胞周期捕获和DNA修复或凋亡(4)。p53在体内许多位点被磷酸化，在体外被许多不同的蛋白激酶磷酸化(5,6)。DNA损伤诱导p53丝氨酸（15、20位点）的磷酸化，从而导致p53与它的负调节子——肿瘤蛋白MDM2的相互作用减少(7)。p38 MAP kinase控制细胞对细胞因子和应激的应答反应(8-11)。在SAPK/JNK通路中，p38 MAP kinase被许多细胞应激激活，包括渗透性休克、炎性细胞因子、脂多糖（LPS）、UV照射和生长因子(8-12)。MKK3, MKK6，SEK通过磷酸化p38 MAP kinase苏氨酸（180位点）和酪氨酸（182位点）而使其活化。生长因子和有丝分裂原诱导 p70 S6 kinase的活化，该激酶又磷酸化S6核糖体蛋白。S6的磷酸化与转位的提高有关，特别是5' 非转位区域内带帽序列的mRNAs(13)。mRNAs (5'TOP)编码的蛋白涉及细胞周期进程和转位机制部分的蛋白，如核糖体蛋白和延长因子(13,14)。

1. Frodin, M. and Gammeltoft, S. (1999) Mol. Cell. Endocrinol. 151, 65-77.
2. Lazar, D.F. et al. (1995) J. Biol. Chem. 270, 20801-20807.
3. Dalby, K.N. et al. (1998) J. Biol. Chem. 273, 1496-1505.
4. Levine, A.J. (1997) Cell 88, 323-331.
5. Meek, D.W. (1994) Semin. Cancer Biol. 5, 203-210.
6. Milczarek, G.J. et al. (1997) Life Sci. 60, 1-11.
7. Shieh, S.Y. et al. (1997) Cell 91, 325-334.
8. Han, J. et al. (1994) Science 265, 808-811.
9. Lee, J.C. et al. (1994) Nature 372, 739-746.
10. Rouse, J. et al. (1994) Cell 78, 1027-1037.
11. Freshney, N.W. et al. (1994) Cell 78, 1039-1049.
12. Raingeaud, J. et al. (1995) J. Biol. Chem. 270, 7420-7426.
13. Peterson, R.T. and Schreiber, S.L. (1998) Curr. Biol. 8, R248-R250.
14. Jefferies, H.B. et al. (1997) EMBO J. 16, 3693-3704.

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 eIF4E antibody detects endogenous levels of its target protein independent of phosphorylation and is provided to control for protein loading.

Source / Purification

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

Description

The PathScan® Multiplex Western Cocktail II 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-p53, phospho-p38 MAPK and phospho-S6 ribosomal protein. The kit also includes elF4E antibody to control protein loading.

Background

The 90 kDa ribosomal S6 kinases (RSK1-3) are a family of serine/threonine kinases broadly expressed in response to many growth factors, polypeptide hormones and neurotransmitters (1). p90RSK is activated by Erk1 and Erk2 in vitro and in vivo via phosphorylation (2). Several sites, such as Ser380, Thr359 and Ser363, are important for its activation (3). 
 The p53 tumor suppressor protein plays a major role in cellular response to DNA damage and other genomic aberrations. Activation of p53 can lead to either cell cycle arrest and DNA repair or apoptosis (4). p53 is phosphorylated at multiple sites in vivo and by several different protein kinases in vitro (5,6). DNA damage induces phosphorylation of p53 at Ser15 and Ser20 and leads to reduced interaction of p53 with its negative regulator, oncoprotein MDM2 (7). 
 p38 MAP kinase controls cellular responses to cytokines and stress (8-11). Like the SAPK/JNK pathway, p38 MAP kinase is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharides (LPS), UV light and growth factors (8-12). MKK3, MKK6 and SEK activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182. 
 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 (13). This group of mRNAs (5'TOP) encodes proteins involved in cell cycle progression and proteins that are part of the translational machinery, such as ribosomal proteins and elongation factors (13,14).

1. Frodin, M. and Gammeltoft, S. (1999) Mol. Cell. Endocrinol. 151, 65-77.
2. Lazar, D.F. et al. (1995) J. Biol. Chem. 270, 20801-20807.
3. Dalby, K.N. et al. (1998) J. Biol. Chem. 273, 1496-1505.
4. Levine, A.J. (1997) Cell 88, 323-331.
5. Meek, D.W. (1994) Semin. Cancer Biol. 5, 203-210.
6. Milczarek, G.J. et al. (1997) Life Sci. 60, 1-11.
7. Shieh, S.Y. et al. (1997) Cell 91, 325-334.
8. Han, J. et al. (1994) Science 265, 808-811.
9. Lee, J.C. et al. (1994) Nature 372, 739-746.
10. Rouse, J. et al. (1994) Cell 78, 1027-1037.
11. Freshney, N.W. et al. (1994) Cell 78, 1039-1049.
12. Raingeaud, J. et al. (1995) J. Biol. Chem. 270, 7420-7426.
13. Peterson, R.T. and Schreiber, S.L. (1998) Curr. Biol. 8, R248-R250.
14. Jefferies, H.B. et al. (1997) EMBO J. 16, 3693-3704.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibody.