p44/42 MAP Kinase Assay Kit (Nonradioactive)

产品编号：CST-9800S 品牌：CST 原厂货号：9800S
产品分类：生化和细胞分析 > 细胞分析试剂盒 > 细胞通路检测试剂盒
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-20°C.

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其它组分：

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (Sepharose® Bead Conjugate) 【子货号：#3510，包装：400 µl，，运保温度：-20°C.】

20X LumiGLO® Reagent and 20X Peroxide【子货号： #7003，包装：5 ml each，，运保温度：-20°C.】

Anti-biotin, HRP-linked Antibody 【子货号：#7075，包装：100 µl，，运保温度：-20°C.】

Anti-mouse IgG, HRP-linked Antibody 【子货号：#7076，包装：100 µl，，运保温度：-20°C.】

Biotinylated Protein Ladder Detection Pack 【子货号：#7727，包装：100 µl，，运保温度：-20°C.】

Elk-1 Fusion Protein 【子货号：#9184，包装：40 µl，，运保温度：-20°C.】

Phospho-Elk-1 (Ser383) (2B1) Mouse mAb 【子货号：#9186，包装：100 µl，，运保温度：-20°C.】

Kinase Buffer (10X) 【子货号：#9802，包装：15 ml，，运保温度：-20°C.】

Cell Lysis Buffer (10X) 【子货号：#9803，包装：15 ml，，运保温度：-20°C.】

ATP (10 mM) 【子货号：#9804，包装：50 µl，，运保温度：-20°C.】

描述：

Nonradioactive p44/42 MAP Kinase Assay Kit提供了检测细胞中p44/42 MAPK激酶活性所需的所有试剂。固定的Phospho-p44/42 MAPK (Thr202/Tyr204) mAb用于免疫沉淀细胞提取物中活化的 p44/42 MAP kinase，接着在体外激酶试验中用Elk-1作为底物。Elk-1的磷酸化由Western blotting分析确定，使用的抗体是Phospho-Elk-1 (Ser383) Antibody。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。

原厂资料：

Description

Nonradioactive p44/42 MAP Kinase Assay Kit provides all the reagents necessary for measuring p44/42 MAP kinase activity in the cell. Immobilized Phospho-p44/42 MAPK (Thr202/Tyr204) mAb is used to immunoprecipitate active p44/42 MAP kinase from cell extracts, then an in vitro kinase assay is performed using Elk-1 protein as a substrate. Elk-1 phosphorylation is then detected by Western blotting using Phospho-Elk-1 (Ser383) Antibody.

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.

注意事项：

For Research Use Only. Not For Use In Diagnostic Procedures.