Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-EGF Receptor (Tyr845) (D63B4) Rabbit mAb #6963

No. Size Price
6963S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
6963 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 175 Rabbit IgG
IP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation,

Homology

Species predicted to react based on 100% sequence homology: Mouse, Rat, Monkey,

Specificity / Sensitivity

Phospho-EGF Receptor (Tyr845) (D63B4) Rabbit mAb recognizes endogenous levels of EGFR protein only when phosphorylated at Tyr845. This antibody may detect other activated ErbB family members. 磷酸化的EGFR (Tyr845)的兔源单克隆抗体(D63B4)仅在Tyr845被磷酸化后才能检测到内源EGFR的水平。本抗体可能与被激活的ErbB家族的其它蛋白发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr845 of human EGFR protein. 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EGFR蛋白Tyr845附近的氨基酸序列合成的。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and A-431 cells, untreated or treated with hEGF #8916 (100 ng/ml for 5 min), using Phospho-EGF Receptor (Tyr845) (D63B4) Rabbit mAb (upper) and EGFR (D38B1) XP® Rabbit mAb #4267 (lower). 用抗磷酸化的EGFR (Tyr845)的兔源单克隆抗体(D63B4)(上)和抗EGFR的兔源单克隆抗体(D38B1) XP® #4267 (下)与下列细胞提取物进行免疫印迹检测:经过或未经hEGF #8916 (100 ng/ml,5分钟) 处理的Hela细胞和A431 细胞的裂解液。

Background

The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, and lysosomal degradation (1,2). Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme, and providing a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of EGFR at Tyr845 (5). The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling (6). Phosphorylation of EGFR at Tyr1045 creates a major docking site for c-Cbl, an adaptor protein that leads to receptor ubiquitination and degradation following EGFR activation (7,8). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (9). A pair of phosphorylated EGFR residues (Tyr1148 and Tyr1173) provides a docking site for the Shc scaffold protein, with both sites involved in MAP kinase signaling activation (2). Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).

表皮生长因子(EGF)受体是一个属于HER/ErbB蛋白家族的跨膜酪氨酸激酶。配体的结合导致了受体的二聚化,自磷酸化,下游信号分子的激活,内化以及溶酶体降解 (1,2)。表皮生长因子受体(EGFR)在激酶域Tyr845位点的磷酸化与稳定激活回路有关,并且维持了酶的激活状态,为底物蛋白提供了可结合的表位(3,4)。c-Src与 EGFR 在 Tyr845位点的磷酸化有关(5)。PLCγ的SH2结构域结合到磷酸化的Tyr992位点,将引起PLCγ介导的下游信号通路的激活(6)。EGFR Tyr1045位点的磷酸化则为c-Cbl生成一个主要的停靠位点,这个衔接蛋白将导致受体的泛素化以及EGFR活化后的降解(7,8)。GRB2衔接蛋白则结合到活化后的EGFR磷酸化的Tyr1068位点(9)。一对磷酸化EGFR残基(Tyr1148 and Tyr1173)为Shc支架蛋白提供了停靠点,这两个位点都与MAPK激酶通路的激活有关(2)。EGFR在特定的丝氨酸和苏氨酸残基上的磷酸化减弱了EGFR激酶的活性。EGFR羧基末端残基Ser1046 和 Ser1047被CaM 激酶II磷酸化,这两个丝氨酸任何一个突变都将导致EGFR酪氨酸自身磷酸化的上调(10)。

  1. Hackel, P.O. et al. (1999) Curr Opin Cell Biol 11, 184-9.
  2. Zwick, E. et al. (1999) Trends Pharmacol Sci 20, 408-12.
  3. Cooper, J.A. and Howell, B. (1993) Cell 73, 1051-4.
  4. Hubbard, S.R. et al. (1994) Nature 372, 746-54.
  5. Biscardi, J.S. et al. (1999) J Biol Chem 274, 8335-43.
  6. Emlet, D.R. et al. (1997) J Biol Chem 272, 4079-86.
  7. Levkowitz, G. et al. (1999) Mol Cell 4, 1029-40.
  8. Ettenberg, S.A. et al. (1999) Oncogene 18, 1855-66.
  9. Rojas, M. et al. (1996) J Biol Chem 271, 27456-61.
  10. Feinmesser, R.L. et al. (1999) J Biol Chem 274, 16168-73.

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

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