PathScan? Total p44/42 MAPK (Erk1/2) Sandwich ELISA Kit

• 产品编号：CST-7050C      品牌：CST       原厂货号：7050C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

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描述：

CST的's PathScan® Total p44/42 MAPK (Erk1/2) Sandwich ELISA Kit #7050能够检测内源性p44/42 MAPK总蛋白水平，如图1所示。该试剂盒的灵敏度如图2所示。该试剂盒经公司内部实验证明能检测已标明物种的蛋白，但也可能能检测其他物种中的类似蛋白。PathScan® Total p44/42 MAPK (Erk1/2) 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p44/p42 MAPK(Erk1/2)总蛋白水平。先将p44/p42 MAPK鼠单抗包被在微孔板上。与细胞裂解物孵育后，p44/42 MAPK便可被包被的抗体捕获。清洗彻底后，加入p44/42 MAPK rabbit detection mAb兔单抗检测p44/42 MAPK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p44/42 MAPK的数量成正比。*试剂盒中抗体是专为本试剂盒特制的。丝裂原活化蛋白激酶(MAPKs)是一个具有广泛保守性的丝氨酸/苏氨酸蛋白激酶家族，在许多细胞过程中起作用，比如细胞增殖、分化、运动和死亡。p44/42 MAPK (Erk1/2)信号转导通路能在应答各种胞外刺激后被激活，这些刺激因子包括有丝分裂原、生长因子和细胞因子(1-3)，并且被研究者认为是诊断和治疗癌症的重要靶点(4)。受到刺激后，三个不同层次的蛋白激酶级联反应就连续被开启，这三个层次的分子包含MAPKKK（MAP3K）、MAPKK（MAP2K）、MAPK。多种p44/42 MAP3Ks已经被鉴定出来，包括Raf家族成员，以及Mos、Tpl2/Cot。MEK1、MEK2是该通路中主要的MAPKKs(5,6)。MEK1、MEK2通过分别磷酸化p44和p42蛋白的活化环内的Thr202/Tyr204位点和Thr185位点/Tyr187位点，从而激活p44、p42。目前已经鉴定出p44/42的一些下游靶点，包括p90RSK (7) 和下游转录因子Elk-1 (8,9)。双特异性(Thr/Tyr) MAPK磷酸酶家族成员DUSPs、MKPs (10)，以及MEK抑制子如U0126、 PD98059，可以负调节p44/42。

1. Roux, P.P. and Blenis, J. (2004) Microbiol Mol Biol Rev 68, 320-44.
2. Baccarini, M. (2005) FEBS Lett 579, 3271-7.
3. Meloche, S. and Pouysségur, J. (2007) Oncogene 26, 3227-39.
4. Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310.
5. Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74.
6. Murphy, L.O. and Blenis, J. (2006) Trends Biochem Sci 31, 268-75.
7. Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
8. Marais, R. et al. (1993) Cell 73, 381-93.
9. Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24.
10. Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Total p44/42 MAPK (Erk1/2) Sandwich ELISA Kit #7050 detects endogenous levels of total p44/42 MAPK protein (see Figure 1). The kit sensitivity is shown in Figure 2. 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® Total p44/42 MAPK (Erk1/2) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of p44/42 MAPK (Erk1/2). A p44/42 MAPK mouse mAb has been coated onto the microwells. After incubation with cell lysates, p44/42 MAPK is captured by the coated antibody. Following extensive washing, a p44/42 MAPK rabbit detection mAb is added to detect the captured p44/42 MAPK protein. Anti-Rabbit IgG, HRP-linked antibody #7074 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of total p44/42 MAPK. 
 Antibodies in kit are custom formulations specific to kit.

Background

Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (Erk1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines (1-3) and is an important target in the diagnosis and treatment of cancer (4). Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family as well as Mos and Tpl2/Cot. MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (10), along with MEK inhibitors such as U0126 and PD98059.

1. Roux, P.P. and Blenis, J. (2004) Microbiol Mol Biol Rev 68, 320-44.
2. Baccarini, M. (2005) FEBS Lett 579, 3271-7.
3. Meloche, S. and Pouysségur, J. (2007) Oncogene 26, 3227-39.
4. Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310.
5. Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74.
6. Murphy, L.O. and Blenis, J. (2006) Trends Biochem Sci 31, 268-75.
7. Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
8. Marais, R. et al. (1993) Cell 73, 381-93.
9. Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24.
10. Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13.

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).