p38 MAP Kinase Assay Kit (Nonradioactive)

• 产品编号：CST-9820S      品牌：CST       原厂货号：9820S
• 产品分类：生化和细胞分析 > 细胞分析试剂盒 > 细胞氧化应激检测试剂盒
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描述：

Nonradioactive p38 MAP Kinase Assay Kit提供所有测量细胞内p38 MAP激酶活性所需的试剂。首先，固定化的Phospho-p38 MAPK (Thr180/Tyr182) mAb用于免疫沉淀p38 MAP激酶，然后以ATF-2为底物进行体内激酶分析。Western blotting分析检测ATF-2 的磷酸化，使用的抗体是Phospho-ATF-2 (Thr71) Antibody。对于人源p38 MAP Kinase苏氨酸（180位点）/酪氨酸（182位点）周围残基，phospho-p38 MAP Kinase (Thr180/Tyr182) mAb通过碳水化合物交联的琼脂糖酰肼珠固定。加入到1X细胞裂解缓冲液和50%甘油中，储存于-20oC，不能分装固定后的抗体。.p38 MAP激酶 (MAPK), 也被称为 RK (1)、CSBP (2)，是哺乳动物体内酵母HOG激酶的同源物，它参与信号传递级联反应，该级联反应能控制细胞对细胞因子和应激反应的应答(1-4)。p38 MAPK的四种异构体, p38α, β, γ (也被称为Erk6、SAPK3), 和δ (也被称为SAPK4)已经被鉴定出来。与SAPK/JNK途径相似，p38 MAPK由各种细胞应激反应激活，包括渗透压休克 、炎性反应、细胞因子、LPS、UV照射和生长因子。(1-5) MKK3, MKK6,SEK磷酸化p38 MAPK蛋白的苏氨酸（180位点）和酪氨酸（182位点）并使其活化。活化的p38 MAPK能将MAPKAP激酶磷酸化、活化 (3)，并磷酸化转录因子ATF-2(5), Max (6), and MEF2 (5-8)。SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole)是p38 MAPK的选择性抑制子。该复合物能通过p38 MAPK抑制MAPKAPK-2的活化和HSP27后续磷酸化 (9)。SB203580通过结合到ATP接合点上而抑制p38 MAPK的摧毁活性，但是不能通过上游激酶而抑制p38 MAPK的磷酸化(10)。

1. Rouse, J. et al. (1994) Cell 78, 1027-37.
2. Han, J. et al. (1994) Science 265, 808-11.
3. Lee, J.C. et al. (1994) Nature 372, 739-46.
4. Freshney, N.W. et al. (1994) Cell 78, 1039-49.
5. Raingeaud, J. et al. (1995) J Biol Chem 270, 7420-6.
6. Zervos, A.S. et al. (1995) Proc Natl Acad Sci U S A 92, 10531-4.
7. Zhao, M. et al. (1999) Mol Cell Biol 19, 21-30.
8. Yang, S.H. et al. (1999) Mol Cell Biol 19, 4028-38.
9. Cuenda, A. et al. (1995) FEBS Lett 364, 229-33.
10. Kumar, S. et al. (1999) Biochem Biophys Res Commun 263, 825-31.

原厂资料：

Description

Nonradioactive p38 MAP Kinase Assay Kit provides all the reagents necessary to measure p38 MAP kinase activity in cells. First, Immobilized Phospho-p38 MAPK (Thr180/Tyr182) mAb is used to immunoprecipitate p38 MAP kinase, then an in vitro kinase assay is performed using ATF-2 as a substrate. ATF-2 phosphorylation is detected by Western blotting using Phospho-ATF-2 (Thr71) Antibody.

Applications

Immobilized Phospho-p38 MAP Kinase (Thr180/Tyr182) AntibodyMonoclonal antibody is produced by immunizing mice with a synthetic phospho-peptide (KLH-coupled) corresponding to residues around Thr180/Tyr182 of human p38 MAP Kinase. The phospho-p38 MAP Kinase (Thr180/Tyr182) mAb was immobilized by crosslinking to agarose hydrazide beads via carbohydrate linkages and are supplied in 1X cell lysis buffer and 50% Glycerol. Store at -20oC. Do not aliquot the immobilized antibody.

Background

p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAP kinase, p38α, β, γ (also known as Erk6 or SAPK3), and δ (also known as SAPK4) have been identified. Similar to the SAPK/JNK pathway, p38 MAP kinase is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharide (LPS), UV light, and growth factors (1-5). MKK3, MKK6, and SEK activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182. Activated p38 MAP kinase has been shown to phosphorylate and activate MAPKAP kinase 2 (3) and to phosphorylate the transcription factors ATF-2 (5), Max (6), and MEF2 (5-8). 
 SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole) is a selective inhibitor of p38 MAPK. This compound inhibits the activation of MAPKAPK-2 by p38 MAPK and subsequent phosphorylation of HSP27 (9). SB203580 inhibits p38 MAPK catalytic activity by binding to the ATP binding pocket, but does not inhibit phosphorylation of p38 MAPK by upstream kinases (10).

1. Rouse, J. et al. (1994) Cell 78, 1027-37.
2. Han, J. et al. (1994) Science 265, 808-11.
3. Lee, J.C. et al. (1994) Nature 372, 739-46.
4. Freshney, N.W. et al. (1994) Cell 78, 1039-49.
5. Raingeaud, J. et al. (1995) J Biol Chem 270, 7420-6.
6. Zervos, A.S. et al. (1995) Proc Natl Acad Sci U S A 92, 10531-4.
7. Zhao, M. et al. (1999) Mol Cell Biol 19, 21-30.
8. Yang, S.H. et al. (1999) Mol Cell Biol 19, 4028-38.
9. Cuenda, A. et al. (1995) FEBS Lett 364, 229-33.
10. Kumar, S. et al. (1999) Biochem Biophys Res Commun 263, 825-31.

注意事项：

For Research Use Only. Not For Use In Diagnostic Procedures