PathScan? Total SAPK/JNK Sandwich ELISA Kit

• 产品编号：CST-7330C      品牌：CST       原厂货号：7330C
• 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：

CST的PathScan® Total SAPK/JNK Sandwich ELISA Kit 能检测内源性总SAPK/JNK蛋白的水平。使用 PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Sandwich ELISA Kit #7325，能检测到经紫外照射处理过的293细胞内phospho-SAPK/JNK被显著诱导了。然而，用PathScan® Total SAPK/JNK Sandwich ELISA Kit #7330检测的总SAPK/JNK 蛋白水平（磷酸化与非磷酸化的）却没有发生改变（图1）。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。CST's PathScan® Total SAPK/JNK Sandwich ELISA Kit是通过固相夹心酶联免疫吸附法（ELISA）来检测内源性的SAPK/JNK蛋白的水平。先将SAPK/JNK (7325-7E7) Mouse mAb*包被在微孔板上。与细胞裂解物孵育后，磷酸化和非磷酸化的SAPK/JNK蛋白便可被包被的抗体捕获。清洗彻底后，加入SAPK/JNK (56G8) Rabbit mAb #9258* 兔单抗检测捕获的SAPK/JNK蛋白的水平。再加入Anti-Rabbit IgG, HRP偶联抗体 #7074*作为二抗来识别之前绑定的检测抗体。最后加入HRP底物—— TMB，进行显色。产生的吸光值与SAPK/JNK蛋白的数量成正比。*试剂盒中抗体是专为本试剂盒特制的。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

CST's PathScan® Total SAPK/JNK Sandwich ELISA Kit detects endogenous levels of total SAPK/JNK protein. Using PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Sandwich ELISA Kit #7325, a significant induction of phospho-SAPK/JNK in 293 cells treated with UV can be detected. However, the level of total SAPK/JNK (phospho and nonphospho), detected by this Sandwich ELISA Kit #7330, remains unchanged (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

CST's PathScan® Total SAPK/JNK Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total SAPK/JNK protein. A SAPK/JNK (7325-7E7) Mouse mAb* has been coated onto the microwells. After incubation with cell lysates, Both nonphospho- and phospho-SAPK/JNK proteins are captured by the coated antibody. Following extensive washing, a SAPK/JNK (56G8) Rabbit mAb #9258* is added to detect the captured SAPK/JNK protein. HRP-linked anti-rabbit antibody #7074* is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total SAPK/JNK protein. 
 * Antibodies in this kit are custom formulations specific to the kit.

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：

* 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).