SAPK/JNK Kinase Assay Kit (Nonradioactive)

• 产品编号：CST-8794S      品牌：CST       原厂货号：8794S
• 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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SAPK/JNK Kinase Assay Kit (Nonradioactive)提供了所有检测细胞中SAPK/JNK激酶活性所需的试剂。连接在琼脂糖珠子上的的phospho-SAPK/JNK Rabbit mAb用于拉下细胞提取物中的SAPK激酶。加入激酶缓冲液，c-Jun融合蛋白和ATP后，SAPK磷酸化c-Jun底物。随后使用Phospho-c-Jun (Ser63)抗体在免疫印迹实验中检测SAPK活性。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)。

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.

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Description

The SAPK/JNK Kinase Assay Kit (Nonradioactive) provides all the reagents necessary to measure SAPK/JNK activity in the cell. A phospho-SAPK/JNK Rabbit mAb linked to agarose beads is used to pull down SAPK enzyme from cell extracts. Upon addition of kinase buffer, c-Jun fusion protein and ATP, SAPK phosphorylates the c-Jun substrate. Phospho-c-Jun (Ser63) Antibody can then be used to measure SAPK activity by immunoblotting.

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, 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：

. Note: Prepare solutions with purified water.