Phospho-p38 MAPK Pathway Sampler Kit

• 产品编号：CST-9913S      品牌：Cell Signaling Technology       原厂货号：9913S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

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描述：

Phospho-p38 MAPK Pathway Sampler Kit试剂盒中的每个抗体都能识别它特异性靶点的唯一磷酸化形式。这些抗体是采用合成的与人源MSK1蛋白Thr581位点、人源 p38 MAPK蛋白Thr180/Tyr182位点、人源MKK3蛋白Ser189/207 位点、人源ATF-2蛋白Thr71位点、人源HSP27蛋白Ser82位点和人源MAPKAPK-2蛋白Thr334位点周围残基相一致的磷酸化肽段免疫动物而获得。多克隆抗体经蛋白A和肽亲和层析纯化。Phospho-p38 MAPK Pathway Sampler Kit提供了一个经济的方法评价p38 MAPK途径中的许多成员的活化状态，包括磷酸化的MSK1, p38 MAPK, MKK3/MKK6, ATF-2, HSP27和 MAPKAPK-2。该试剂盒包含足够的一抗、二抗完成四个Wester blot实验。p38 MAP kinase (MAPK),也被称为RK (1)或CSBP (2)，是酵母中HOG激酶在哺乳动物中的同源蛋白，在细胞因子和应急诱导的信号转导过程中起重要作用。(1-4)。已经发现了4种亚型的p38 MAPK，分别是p38α, β, γ（也被称为Erk6或 SAPK3）以及δ (也被称为 SAPK4)。与SAPK/JNK通路类似，p38 MAPK可以被多种细胞压力所激活，包括渗透压休克，炎症因子，脂多糖（LPS），紫外线照射和生长因子（1-5）。MKK3, MKK6和SEK能够使p38 MAPK的Thr180 和Tyr182位点磷酸化，从而激活p38 MAPK。激活的p38 MAPK能够磷酸化并激活MAPKAP kinase 2 (3)，进而最终磷酸化ATF-2 (5), Max (6)和MEF2等转录因子(5-8)。SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole)是p38 MAPK的选择性抑制剂。该复合物通过抑制p38 MAPK及后续HSP27的磷酸化，抑制MAPKAPK-2的激活（9）。SB203580能够结合到p38 MAPK与ATP结合的位置从而抑制p38 MAPK的催化活性，但不会通过上游激酶抑制p38 MAPK的磷酸化（10）。在体外激酶实验中，MAPKAPK-2的四个残基（Thr25、Thr222、Ser272和Thr334）能够被p38磷酸化(3)。Thr222、Ser272和Thr334位点的磷酸化似乎对MAPKAPK-2 的活化十分重要(3)。活化的MAPKAPK-2蛋白又能将HSP27蛋白的Ser15、Ser78和Ser82位点磷酸化 (3,9)。HSP27的磷酸化能够引起其三级结构的改变，从大的同型多聚体变为二聚体、单体(10)。实验证明，HSP27的磷酸化和浓度的增加会调节肌动蛋白的聚合作用和重组(11,12)。

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.
11. Landry, . et al. (1992) J. Biol. Chem. 267, 794-803.
12. Rogalla, . et al. (1999) J. Biol. Chem. 274, 18947-18956.
13. Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
14. Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.

原厂资料：

Specificity / Sensitivity

Each antibody in the Phospho-p38 MAPK Pathway Sampler Kit recognizes only the phosphorylated form of its specific target.

Source / Purification

Antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Thr581 of human MSK1, Thr180/Tyr182 of human p38 MAPK, Ser189/207 of human MKK3, Thr71 of human ATF-2, Ser82 of human HSP27 or Thr334 of human MAPKAPK-2. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Description

The Phospho-p38 MAPK Pathway Sampler Kit provides an economical means to evaluate the activation status of multiple members of the p38 MAPK pathway, including phosphorylated MSK1, p38 MAPK, MKK3/MKK6, ATF-2, HSP27 and MAPKAPK-2. The kit includes enough primary and secondary antibodies to perform four Western blot experiments.

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).Four residues (Thr25, Thr222, Ser272 and Thr334) of MAPKAPK-2 are phosphorylated by p38 in an in vitro kinase assay (3). Phosphorylation at Thr222, Ser272 and Thr334 seems to be essential for the activity of MAPKAPK-2 (3). Activated MAPKAPK-2 can in return phosphorylate HSP27 at serines 15, 78 and 82 (3,9). Phosphorylation of HSP27 causes a change in the tertiary structure of HSP27, which shifts from large homotypic multimers to dimmers and monomers (10). It has been illustrated that phosphorylation and increased concentration of HSP27 modulate actin polymerization and reorganization (11,12).

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.
11. Landry, . et al. (1992) J. Biol. Chem. 267, 794-803.
12. Rogalla, . et al. (1999) J. Biol. Chem. 274, 18947-18956.
13. Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
14. Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.

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