PhosphoPlus? SAPK/JNK (Thr183/Tyr185) Antibody Kit

• 产品编号：CST-9250S      品牌：CST       原厂货号：9250S
• 产品分类：生化和细胞分析 > 细胞分析试剂盒 > 细胞通路检测试剂盒
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描述：

Phospho-SAPK/JNK (Thr183/Tyr185) Antibody兔多抗能检测所有三种SAPK/JNK亚型的双磷酸化蛋白水平。Western分析表明该抗体不能与内源性p44/42 MAPK或 p38 MAPK的磷酸化形式发生交叉反应。然而，由于SAPK/JNK和 p44/42 MAP激酶活化位点的近似同源性，大量的 phospho-p44/42 MAPK可能会被检测到。该多克隆抗体应激活化蛋白激酶/Jun氨基末端激酶SAPK/JNK能够有效、优先地被多种环境压力激活，这些环境压力包括紫外照射和gamma 辐射、神经酰胺、炎性因子，在某些情况下也包括生长因子和GPCR激动剂 (1-6)。正如其他的 MAPKs，核心信号传导单元包括一个MAPKKK，通常是MEKK1-MEKK4，或一种混合谱系酶 (MLKs），这些MAPKKK激酶能磷酸化并激活 MKK4/7。活化后，MKKs磷酸化并激活SAPK/JNK 激酶 (2)。应激反应信号经Rho 家族(Rac, Rho, cdc42)的小GTP酶被传递到这个级联信号反应中 (3)。Rac1 和 cdc42蛋白都能调节MEKKs、 MLKs的激活(3)。另外一种情况是，MKK4/7能够通过生发中心激酶 (GCK)家族成员的刺激，在一个非GTP酶依赖机制中被激活 (4)。有三种不同的SAPK/JNK基因，而每一种都会经过选择性剪切产生多种异构体(3)。活化的二聚体SAPK/JNK，能转位到细胞核中，通过它对 c-Jun, ATF-2和其他转录因子的效应而调节转录过程(3,5)。是采用合成的与SAPK/JNK蛋白Thr183/Tyr185周围残基相一致的磷酸化肽段免疫动物而获得。该抗体经蛋白A和肽亲和层析纯化。

1. Davis, R.J. (1999) Biochem Soc Symp 64, 1-12.
2. Ichijo, H. (1999) Oncogene 18, 6087-93.
3. Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69.
4. Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62.
5. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
6. Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem Sci 23, 481-5.

原厂资料：

Specificity / Sensitivity

Phospho-SAPK/JNK (Thr183/Tyr185) Antibody detects the dually phosphorylated isoforms of all three SAPKs/JNKs. Western analysis shows that this antibody does not cross-react with endogenous levels of the corresponding phosphorylated forms of p44/42 MAP Kinase or p38 MAP Kinase. However, because of the close homology between the active sites of SAPK/JNK and p44/42 MAP Kinase, larger amounts of phospho-p44/42 MAP Kinase may be detectable.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr183/Tyr185 of human SAPK/JNK. Antibodies are purified by protein A and peptide affinity chromatography.Monoclonal antibody is produced by immunizing animals with a human JNK2/MBP fusion protein.

Description

The PhosphoPlus® SAPK/JNK (Thr183/Tyr185) Antibody Kit provides reagents and protocols for the rapid analysis of SAPK/JNK phosphorylation. 
 SAPK/JNK Control Cell Extracts: Total cell extracts from 293 cells prepared with and without UV light treatment serve as positive and negative controls.

Background

The stress-activated protein kinase/Jun-amino-terminal kinase SAPK/JNK is potently and preferentially activated by a variety of environmental stresses including UV and gamma radiation, ceramides, inflammatory cytokines, and in some instances, by growth factors and GPCR agonists (1-6). As with the other MAPKs, the core signaling unit is composed of a MAPKKK, typically MEKK1-MEKK4, or by one of the mixed lineage kinases (MLKs), which phosphorylate and activate MKK4/7. Upon activation, MKKs phosphorylate and activate the SAPK/JNK kinase (2). Stress signals are delivered to this cascade by small GTPases of the Rho family (Rac, Rho, cdc42) (3). Both Rac1 and cdc42 mediate the stimulation of MEKKs and MLKs (3). Alternatively, MKK4/7 can be activated in a GTPase-independent mechanism via stimulation of a germinal center kinase (GCK) family member (4). There are three SAPK/JNK genes each of which undergoes alternative splicing resulting in numerous isoforms (3). SAPK/JNK, when active as a dimer, can translocate to the nucleus and regulate transcription through its effects on c-Jun, ATF-2, and other transcription factors (3,5).

1. Davis, R.J. (1999) Biochem Soc Symp 64, 1-12.
2. Ichijo, H. (1999) Oncogene 18, 6087-93.
3. Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69.
4. Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62.
5. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
6. Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem Sci 23, 481-5.

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 antibodies