Cell Signaling Technology

Product Pathways - Metabolism

Phospho-PKM2 (Tyr105) Antibody #3827

No. Size Price
3827S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
3827 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 60 Rabbit

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting,

Specificity / Sensitivity

Phospho-PKM2 (Tyr105) Antibody detects endogenous levels of PKM2 protein only when phosphorylated at Tyr105. This antibody may slightly cross react with PKM1 phosphorylated at the equivalent site.

Phospho-PKM2 (Tyr105)抗体识别内源性的Tyr105磷酸化的PKM2蛋白水平。它也可以稍弱地识别对应部位磷酸化的PKM1蛋白。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to the sequence surrounding Tyr105 of human PKM2 protein. Antibodies are purified by protein A and peptide affinity chromatography.

该多克隆抗体用与人类PKM2中Tyr105附近的氨基酸序列对应的人工合成磷酸化肽段免疫动物而制成。该抗体使用蛋白A和肽亲和层析纯化而得。

Western Blotting

Western Blotting

Western blot analysis of extracts from A549 and DU 145 cells using Phospho-PKM2 (Tyr105) Antibody (upper) or PKM2 Antibody #3198 (lower).

对A549细胞和DU 145细胞抽提液,使用Phospho-PKM2 (Tyr105)抗体(上图)或PKM2抗体#3198(下图)进行Western blot分析。

Western Blotting

Western Blotting

GST-PKM2 wild-type or the Tyr105Phe mutant was incubated in an in vitro kinase assay in the presence or absence of active FGFR1. Western blot analysis was performed using Phospho-PKM2 (Tyr105) Antibody and a phospho-Tyr antibody. The data demonstrate the specificity of the Phospho-PKM2 (Tyr105) Antibody and that the Tyr105Phe mutation abolishes PKM2 phosphorylation at Tyr105 by FGFR1 in vitro. (Adapted from Hitosugi, T. et al., 2009).

GST-PKM2野生型或Tyr105Phe突变体在体外孵育以在活性FGFR1存在或不存在时进行激酶测试。使用Phospho-PKM2 (Tyr105)抗体和phospho-Tyr抗体进行Western blot分析。数据显示了Phospho-PKM2 (Tyr105)的特异性,Tyr105Phe突变消除了PKM2被FGFR1在体外对其Tyr105的磷酸化。(修改自Hitosugi, T. et al., 2009)

Western Blotting

Western Blotting

Western blot analysis of NCI-H1299 cells using Phospho-PKM2 (Tyr105) Antibody, total PKM2 Antibody #3198, total FGFR1 antibody, phospho-Tyr antibody, and β-actin antibody. The data demonstrate that inhibition of FGFR1 by TKI258 treatment in NCI-H1299 cells results in decreased Tyr105 phosphorylation of endogenous PKM2. (Adapted from Hitosugi, T. et al., 2009).

使用Phospho-PKM2 (Tyr105)抗体,总PKM2抗体#3198,总FGFR1抗体,phospho-Tyr抗体和β-actin抗体对NCI-H1299细胞进行Western blot分析。数据显示TKI258处理NCI-H1299细胞抑制FGFR1,导致内源性PKM2 Tyr105的磷酸化降低。(修改自Hitosugi, T. et al., 2009)

Background

Pyruvate kinase, a glycolytic enzyme, catalyses the conversion of phosphoenolpyruvate to pyruvate. In mammals, the M1 isoform (PKM1) is expressed in most adult tissues (1). The M2 isoform (PKM2), an alternatively-spliced variant of M1, is expressed during embryonic development (1). Studies found that cancer cells exclusively express PKM2 (1-3). PKM2 is shown to be essential for aerobic glycolysis in tumors (Warburg effect) (1). When the M2 isoform is switched to the M1 isoform, aerobic glycolysis is reduced and oxidative phosphorylation is increased in cancer cells (1). These cells also show decreased tumorigenicity in mouse xenografts (1). Recent studies show that the oncogenic forms of FGFR1 directly phosphorylate Tyr105 of PKM2 and thereby inhibit the formation of active tetrameric PKM2 (4). A PKM2 mutant found in cancer cells, in which Tyr105 is replaced by phenylalanine, leads to reduced cell proliferation in hypoxia and tumor growth in xenografts in nude mice (4). These findings suggest that the phosphorylation at Tyr105 is a critical switch for the metabolism in cancer cells that promotes tumor growth (4).

丙酮酸激酶是一种糖酵解酶,催化丙酮酸转变为磷酸丙酮酸。在哺乳动物中,M1亚型(PKM1)在大多数成年组织中表达(1)。M2亚型(PKM2)是M1亚型的一种可变剪切结果,在胚胎发育时表达(1)。研究发现癌细胞只表达PKM2(1-3)。PKM2对肿瘤的无氧糖酵解(Warburg效应)很关键(1)。当M2亚型转变为M1亚型时,癌细胞无氧酵解下降,氧化磷酸化增加(1)。在鼠异源移植时,这些细胞也显示出降低的成瘤可能(1)。最近的研究表明FGFR1的致癌形式直接磷酸化PKM2的Tyr105,从而抑制PKM2活性四聚体的形成(4)。一种PKM2突变在癌细胞中发现,Tyr105被苯丙氨酸代替,导致在低氧环境下细胞增生的减低和裸鼠上异源移植的肿瘤生长变缓(4)。这些发现表明Tyr105的磷酸化对癌细胞生长所需的代谢转变十分关键(4)。

Phosphorylation of PKM2 on Tyr105 was identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's LC-MS/MS platform for phosphorylation site discovery.

PKM2的Tyr105的磷酸化位点由CST公司使用PhosphoScan和CST的磷酸化位点液相色谱-质谱/质谱平台发现。

  1. Christofk, H.R. et al. (2008) Nature 452, 230-3.
  2. Mazurek, S. et al. (2005) Semin Cancer Biol 15, 300-8.
  3. Dombrauckas, J.D. et al. (2005) Biochemistry 44, 9417-29.
  4. Israelsen, W.J. et al. (2013) Cell 155, 397-409.
  5. Hitosugi, T. et al. (2009) Sci Signal 2, ra73.

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

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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