Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody #9101

ERK   ERK1   erk1/2   ERK2   mapk   p42   p44   p44/42   pmapk   sc-16982   sc-7976  

No. Size Price
9101L 600 µl ( 60 western blots ) ¥8,792.00 现货查询 购买询价
9101S 200 µl ( 20 western blots ) ¥3,780.00 现货查询 购买询价
9101 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Hamster,Monkey,Mink,D. melanogaster,Zebrafish,Bovine,Pig,C. elegans, Endogenous 42, 44 Rabbit
IP 1:50
F 1:200
IF-IC 1:250

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Homology

Species predicted to react based on 100% sequence homology: Chicken,

Specificity / Sensitivity

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody detects endogenous levels of p44 and p42 MAP Kinase (Erk1 and Erk2) when phosphorylated either individually or dually at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2). The antibody does not cross-react with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP Kinase, and does not cross-react with non-phosphorylated Erk1/2.

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody兔多抗能够检测内源性的Thr202和Tyr204被同时或单独磷酸化的Erk1(Thr185和 Tyr187被同时或单独磷酸化的Erk2)。该抗体与JNK/SAPK或p38 MAPK中的相应磷酸化残基没有交叉反应,与非磷酸化的Erk1/2没有交叉反应。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase. Antibodies are purified by protein A and peptide affinity chromatography.

该多克隆抗体是采用合成的与人源 p44 MAPK的Thr202位点/Tyr204位点周围序列相一致的磷酸化肽段免疫动物后产生的。

Western Blotting

Western Blotting

Specificity and sensitivity of Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody. The antibody reacts specifically with as little as 0.25 ng of phosphorylated p42 MAP kinase and does not cross-react with up to 4 µg of nonphosphorylated p42 MAP kinase.

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody兔多抗的特异性和灵敏度:该抗体对磷酸化的p42 MAPK的检测灵敏度可达0.25ng,对非磷酸化的p42 MAPK在多达4ug时也不会产生非特异结合。

Flow Cytometry

Flow Cytometry

Phosphorylated MEK and Erk were assayed in human peripheral blood lymphocytes stimulated with PMA in the presence or absence of the Raf inhibitor BAY 37-9751 or the MEK inhibitor U0126 #9903. BAY 37-951 blocked PMA-stimulated phosphorylation of both MEK and Erk, consistent with inhibition at the level of Raf, while U0126 blocked phosphorylation of Erk only, consistent with inhibition at the level of MEK. From Chow, S. et al. (2001) Cytometry 46, 72-78.

在Raf抑制剂BAY 37-9751或MEK抑制剂U0126#9903存在的情况下,在经PMA刺激的人上皮血液淋巴细胞中用流式方法检测磷酸化的MEK、Erk。BAY 37-951阻断了PMA刺激的MEK、Erk两者的磷酸化,并与Raf水平抑制的结果相一致。而U0126只阻断Erk的磷酸化,并与MEK水平的抑制相一致。来自于Chow, S. 等 (2001) Cytometry 46, 72-78.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green) or PMA-treated (blue), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody compared to a nonspecific negative control antibody (red).

流式方法检测细胞提取物:未经处理的 (蓝色)、PMA处理的(绿色)Jurkat细胞,使用的抗体是Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody兔多抗,红色曲线表示使用非特异性阴性对照抗体的结果。

Western Blotting

Western Blotting

Western blot analysis of whole-cell extracts from unstarved wild-type mouse embryonic fibroblasts (MEFs) treated with the indicated combinations of basic Fibroblast Growth Factor (bFGF #9952, 100 ng/ml for 30 minutes), Platelet-Derived Growth Factor (PDGF #9909, 100 ng/ml for 30 minutes), MEK1 Inhibitor (PD98059 #9900, 50 µM, 2 hour pre-treatment), and MEK1/2 Inhibitor (U0126 #9903, 10 µM, 2 hour pre-treatment), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody #9101 (upper panel) and p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower panel).

Western blot方法检测细胞提取物:基础成纤维细胞生长因子(bFGF #9952, 100 ng/ml 30 min)、血小板衍生生长因子(PDGF #9909, 100 ng/ml 30 min)、MEK1抑制剂(PD98059 #9900, 50 µM, 提前处理2 h),和MEK1/2抑制剂(U0126 #9903, 10 µM, 提前处理2 h)的不同组合处理的非饥饿野生型小鼠胚胎成纤维细胞(MEFs),使用的抗体是Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody #9101兔多抗 (上图)和p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695兔单抗 (下图)。

IF-IC

IF-IC

Confocal immunofluorescent analysis of NIH/3T3 cells either U0126-treated (left) or PDGF-treated (right) and labeled with Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

激光共聚焦荧光法检测NIH/3T3细胞,左侧细胞用U0126处理,右侧细胞用PDGF处理并用Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody兔多抗标记(呈绿色)。肌动蛋白纤维丝用Alexa Fluor® 555鬼笔环肽标记(呈红色)。

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.

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.

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