PathScan? Phospho-SAPK/JNK (Thr183/Tyr185) Chemiluminescent Sandwich ELISA Kit

• 产品编号：CST-7849C      品牌：CST       原厂货号：7849C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Chemiluminescent Sandwich ELISA Kit #7849能检测人和小鼠内源性phospho-SAPK/JNK (Thr183/Tyr185)蛋白的水平。经内部测试确认，本试剂盒可以检测到所列物种中的蛋白。但对其他物种中的同源蛋白可能也可以检测。PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Chemiluminescent Sandwich ELISA Kit是通过固相夹心酶联免疫吸附法（ELISA），用化学发光读数来测定内源性Phospho-SAPK/JNK (Thr183/Tyr185) 蛋白水平。与常规的显色检测相比，化学发光ELISAs 通常具有更广的动态范围和更高的灵敏度。该化学发光ELISA所提供的微孔板为低容量孔板，当使用小体积的样本量时，显示出增强的信号和灵敏度。先将SAPK/JNK mouse mAb鼠单抗包被在微孔板上。与细胞裂解物孵育后，包括磷酸化和非磷酸化的SAPK/JNK总蛋白便可被包被的抗体捕获。清洗彻底后，加入phospho-SAPK/JNK (Thr183/Tyr185) antibody兔多抗检测被捕获的phospho-SAPK/JNK (Thr183/Tyr185)蛋白。再加入Anti-rabbit IgG, HRP偶联抗体作为二抗来识别之前绑定的检测抗体。最后加入化学发光试剂，用于信号显示。产生的相对光单位的数值与phospho-SAPK/JNK (Thr183/Tyr185) 蛋白的数量成正比。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.

原厂资料：

Specificity / Sensitivity

PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Chemiluminescent Sandwich ELISA Kit #7849 detects endogenous levels of phospho-SAPK/JNK (Thr183/Tyr185) in human and mouse cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-SAPK/JNK (Thr183/Tyr185) protein with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. A SAPK/JNK mouse mAb has been coated on the microwells. After incubation with cell lysates, total SAPK/JNK protein (non-phosphorylated and phosphorylated) is captured by the coated antibody. Following extensive washing, phospho-SAPK/JNK (Thr183/Tyr185) antibody is added to detect the captured phospho-SAPK/JNK (Thr183/Tyr185) protein. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-SAPK/JNK (Thr183/Tyr185) protein.

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：

Low volume microplate * 12 8-well modules -each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).