Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb #3077

c-met  

No. Size Price
3077S 100 µl ( 10 western blots ) ¥4,050.00 现货查询 购买询价
3077T 20 µl ( 2 western blots ) ¥1,600.00 现货查询 购买询价
3077 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 145 Rabbit
IP 1:50
IHC-P 1:320
F 1:200
IF-IC 1:800
IHC-F 1:320

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), IHC-F=Immunohistochemistry (Frozen),

Specificity / Sensitivity

Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb detects endogenous levels of Met only when phosphorylated at Tyr1234/1235. This antibody may cross-react with overexpressed tyrosine phosphorylated Src proteins in Western blot.

磷酸化Met (Tyr1234/1235) (D26) XP® 兔单抗仅在Tyr1234/1235位点磷酸化时能检测到内源的Met蛋白的表达。在免疫印迹检测中本抗体可能其它过表达磷酸化酪氨酸的Src蛋白发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1234/1235 of human Met.

单克隆抗体通过用多肽免疫动物得到,该磷酸化多肽是根据人的Met蛋白Tyr1234/1235附近的氨基酸序列合成的。

Western Blotting

Western Blotting

Western blot analysis of cell extracts from HeLa cells, untreated or stimulated with HGF, using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (upper) and Met (25H2) Mouse mAb #3127 (lower).用Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (上) 和 Met (25H2) Mouse mAb #3127 (下)对未处理的和HGF处理的Hela细胞的提取物进行免疫印迹检测。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb.用Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb对石蜡包埋的人肺癌组织进行免疫组化检测:左,未经处理;右,λ phosphatase处理。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis on Src-transfected NIH/3T3 cells, using a Phospho-Src Family (Tyr416) Antibody (left) or Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (right), indicating that the antibody does not cross-react with Src phosphorylated at Tyr416 via immunohistochemistry.用Phospho-Src Family (Tyr416) Antibody (左) 和Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb(右)对转染了Src的NIH/3T3细胞进行免疫组化检测,结果显示本抗体在免疫组化中不与Tyr416磷酸化的Src发生交叉反应。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohisochemical analysis of paraffin-embedded HCC827 xenograft using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb. 用Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb对石蜡包埋的HCC827细胞异种移植物进行免疫组化检测。

IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen MKN45 xenograft using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb.用Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb对MKN45异种移植物的冰冻组织进行免疫组化检测。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded xenografts from 3T3-Met (left) and 3T3-Ron cells (right) using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb, indicating that this antibody does not cross-react with activated Ron by immunohistochemistry. Image courtesy of Pfizer, Inc. 用Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb对石蜡包埋的3T3-Met (左) 和 3T3-Ron(右)细胞的异种移植物进行免疫组化检测,结果显示本抗体在免疫组化中不与激活的Ron发生交叉反应。图片来自Pfizer, Inc。

Western Blotting

Western Blotting

Western blot analysis of purified active Ron kinase using a Phospho-Ron (Ser1349) Antibody (A), a Phospho-Ron (Tyr1238) Antibody (B), Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (C) and Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 (D). This demonstrates that Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb does not cross-react with phospho-Ron by western analysis.用Phospho-Ron (Ser1349) Antibody (A), Phospho-Ron (Tyr1238) Antibody (B), Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (C) 和 Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 (D)对纯化的激活态Ron激酶进行免疫印迹检测,结果显示免疫印迹检测中Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb不与 phospho-Ron发生交叉反应。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded papillary renal cell carcinoma using Phospho-Met (Tyr1234/1225) (D26) XP® Rabbit mAb.用Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb对石蜡包埋的人乳突状肾细胞癌组织进行免疫组化检测。

IF-IC

IF-IC

Confocal immunofluorescence analysis of MKN45 cells, untreated (left) or treated with SU11274 (1 μM, 3 hours; right), using Phospho-MET (Tyr1234/Tyr1235) (D26) XP® Rabbit mAb. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).激光共聚焦荧光法检测MKN45细胞:未处理的(左)和SU11274(1 μM, 3 hours) 处理的(右)细胞,使用抗体为:Phospho-MET (Tyr1234/Tyr1235) (D26) XP® Rabbit mAb。蓝色伪彩为DNA荧光染料(产品信息为 DRAQ5®#4084 )。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of MKN-45 cells, untreated (green) or treated with SU11274 (blue).对MKN-45细胞进行流式细胞术检测:未处理的(绿色),SU11274 处理的(蓝色)。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb on SignalSlide™ Phospho-Met (1234/1235) IHC Controls #8118 [MKN45 cells, untreated (left) or SU11274-treated (right)].

Background

Met, a high affinity tyrosine kinase receptor for hepatocyte growth factor (HGF, also known as scatter factor) is a disulfide-linked heterodimer made of 45 kDa α- and 145 kDa β-subunits (1,2). The α-subunit and the amino-terminal region of the β-subunit form the extracellular domain. The remainder of the β-chain spans the plasma membrane and contains a cytoplasmic region with tyrosine kinase activity. Interaction of Met with HGF results in autophosphorylation at multiple tyrosines, which recruit several downstream signaling components, including Gab1, c-Cbl, and PI3 kinase (3). These fundamental events are important for all of the biological functions involving Met kinase activity. The addition of a phosphate at cytoplasmic Tyr1003 is essential for Met protein ubiquitination and degradation (4). Phosphorylation at Tyr1234/1235 in the Met kinase domain is critical for kinase activation. Phosphorylation at Tyr1349 in the Met cytoplasmic domain provides a direct binding site for Gab1 (5). Altered Met levels and/or tyrosine kinase activities are found in several types of tumors, including renal, colon, and breast. Thus, Met is an attractive cancer therapeutic and diagnostic target (6,7).

Met作为肝细胞生长因子 (HGF, 亦作扩散因子) 的高亲和性酪氨酸激酶受体是一个二硫键连接的异二聚体,含有一个45 kDa的 α-亚基和一个145 kDa的 β-亚基 (1,2)。α-亚基和β-亚基的氨基端形成胞外区,β-亚基的其余部分跨细胞膜并且含有一个具有激酶活性的胞内区。Met和HGF相互作用会导致多个酪氨酸位点的自磷酸化,继而招募下游信号分子Gab1, c-Cbl和 PI3 K激酶(3)。这一系列基础性事件对于Met激酶相关的所有生物学功能十分重要。胞内Tyr1003位点的磷酸化对于Met的泛素化和降解是必须的(4)。激酶结构域Tyr1234/1235的磷酸化对于Met的激酶活性十分关键。胞内 Tyr1349的磷酸化为Met提供了Gab1 的直接结合位点(5)。在肾、结肠和乳腺癌中发现有Met水平和/或激酶活性的改变,因此Met是肿瘤治疗和诊断的一个靶点(6,7)。

  1. Cooper, C.S. et al. (1984) Nature 311, 29-33.
  2. Bottaro, D.P. et al. (1991) Science 251, 802-4.
  3. Bardelli, A. et al. (1997) Oncogene 15, 3103-11.
  4. Taher, T.E. et al. (2002) J Immunol 169, 3793-800.
  5. Schaeper, U. et al. (2000) J Cell Biol 149, 1419-32.
  6. Eder, J.P. et al. (2009) Clin Cancer Res 15, 2207-14.
  7. Sattler, M. and Salgia, R. (2009) Update Cancer Ther 3, 109-118.

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