Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370

ERK   ERK 1/2   erk-1   erk-2   mapk   p42   p44   p44/42   p44/42 MAPK   pmapk   sc-16982   sc-7976  

No. Size Price
4370L 600 µl ( 120 western blots ) ¥9,448.00 现货查询 购买询价
4370S 200 µl ( 40 western blots ) ¥3,990.00 现货查询 购买询价
4370T 20 µl ( 2 western blots ) ¥1,600.00 现货查询 购买询价
4370 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:2000 Human,Mouse,Rat,Hamster,Monkey,Mink,D. melanogaster,Zebrafish,Bovine,Dog,Pig,S. cerevisiae, Endogenous 44, 42 Rabbit IgG
IP 1:50
IHC-P 1:400
F 1:800
IF-IC 1:200

Species cross-reactivity is determined by western blot.

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

Homology

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

Specificity / Sensitivity

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb 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.

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb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。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase.

该单克隆抗体是采用合成的与人源p44 MAPK的Thr202/Tyr204位点周围残基一致的磷酸化肽段免疫动物而产生的。

Western Blotting

Western Blotting

Western blot analysis of extracts from 293, NIH/3T3 and C6 cells, treated with λ phosphatase or TPA as indicated, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (upper), or p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb on SignalSlide™ Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Controls #8103 (paraffin-embedded NIH/3T3 cells, treated with U0126 #9903 (left) or TPA #4174 (right).

免疫组织化学方法检测SignalSlide™ Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Controls #8103(石蜡包埋的NIH/3T3细胞,左侧为经U0126 #9903处理,右侧为经TPA #4174 处理)。使用的抗体为中的Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb兔单抗。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.

免疫组织化学方法检测石蜡包埋的人肺癌组织,左侧样品未经处理,右侧样品经λ磷酸酶处理。使用的抗体为Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb兔单抗。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.

免疫组织化学方法检测石蜡包埋的人乳腺癌组织。使用的抗体为Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb兔单抗。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, U0126-treated (blue) or PMA-treated (green), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.

流式方法检测细胞提取物:U0126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) (D13.14.4E) XP® Rabbit mAb兔单抗。

IF-IC

IF-IC

Confocal immunofluorescent analysis of C2C12 cells, treated with U0126 #9903 (left, 10 μM for 1 h) or TPA #9905 (right, 200 nM for 15 min), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

激光共聚焦荧光法检测C2C12细胞,左侧细胞用U0126 #9903 (10 μM 1 h)处理,右侧细胞用U0126 #9903 (10 μM 1 h)处理。使用的抗体是Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb兔单抗(呈绿色)。激动蛋白纤维丝用Alexa Fluor® 555鬼笔环肽标记(呈红色)。蓝色伪彩为DNA荧光染料(产品信息为 DRAQ5®#4084)。

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untreated or treated with either U0126 #9903 (10 µM for 1h) or TPA #9905 (200 nM for 10 m), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 and p44/42 MAPK (Erk1/2) (3A7) Mouse mAb #9107.

IF-IC

IF-IC

Confocal immunofluorescent analysis of Drosophila egg chambers, untreated (top) or λ phosphatase-treated (bottom), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

激光共聚焦荧光法检测卵室,上侧细胞未经处理,下侧细胞用λ磷酸酶处理。使用的抗体是Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370兔单抗(呈绿色)和S6 Ribosomal Protein (54D2) Mouse mAb #2317鼠单抗(呈红色)。蓝色伪彩为DNA荧光染料(产品信息为 DRAQ5®#4084)。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HT1080 cells, starved overnight then treated with U0126 #9903 (10 uM, 2 h; left) or PDBu (Phorbol 12,13-Dibutyrate) #12808 (100 nM, 15 m; right) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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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