PhosphoPlus? p44/42 MAPK (Erk1/2)(Thr202/Tyr204) In-Cell Duet (ICW Compatible)

• 产品编号：CST-7263L      品牌：CST       原厂货号：7263L
• 产品分类：抗体 > 一抗 > 抗体对
• 应用分类：

其它组分：

描述：

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204)兔单抗能够检测内源性的Thr202和Tyr204被双重磷酸化的Erk1(Thr185和 Tyr187双重磷酸化的Erk2)，以及Thr202被单独磷酸化的p44和p42 MAPK(Erk1和Erk2)蛋白。该抗体与JNK/SAPK或p38 MAPK的相应磷酸化残基没有交叉反应。Total p44/42 MAPK能检测内源性总p44 、 p42 MAP(Erk1和 Erk2)蛋白水平。在一些系统中与p44/Erk1相比，该抗体可能更容易识别p42/Erk2 。该抗体不能与SAPK/JNK 、p38 MAP激酶发生交叉反应。该单克隆抗体是采用合成的与人源p44 MAPK Thr202/Tyr204位点（或p42 MAPK）周围残基相一致的磷酸化肽段（或肽段）免疫动物而获得。Cell Signaling Technology (CST)的PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) In-Cell Duet为评估蛋白活性状态提供了一个简单方法，使用了具有近红外检测能力的多孔板扫描技术，如LI-COR® Biosciences Odyssey® Infrared Imaging System。该试剂盒包含预先优化的活化状态蛋白抗体和总蛋白抗体的混合物，这两种抗体精选自CST产品，具有卓越的性能。磷酸化蛋白和总蛋白在相同的微孔中进行同时检测，使磷酸化蛋白的水平可以根据总蛋白的量进行标准化。试剂盒中还包括了近红外检测cocktail。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

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) detects endogenous levels of p44 and p42 MAP kinase (Erk1 and Erk2) when dually phosphorylated at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2), and singly phosphorylated at Thr202. This antibody does not cross-react with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP kinases. Total p44/42 MAPK detects endogenous levels of total p44/42 MAP kinase (Erk1/Erk2) protein. In some systems this antibody may recognize p42/Erk2 more readily than p44/Erk1. The antibody does not cross-react with JNK/SAPK or p38 MAP kinase.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase or to the sequence of p42 MAP kinase.

Description

PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) In-Cell Duet from Cell Signaling Technology (CST) provides an easy method to assess protein activation status using a multi-well plate scanner with near infrared detection capabilities, such as the LI-COR® Biosciences Odyssey® Infrared Imaging System. This kit contains a pre-optimized activation-state and total protein antibody cocktail, selected based on superior performance. Phosphorylated and total protein are detected simultaneously in the same well, allowing levels of phosphorylated protein to be normalized to total protein. A near infrared detection cocktail is also included.

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：

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 either cocktail.