Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb #8869

No. Size Price
8869S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
8869T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
8869 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 120 Rabbit IgG
IP 1:50

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,

Specificity / Sensitivity

Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb recognizes endogenous levels of c-Cbl protein only when phosphorylated at Tyr700.

Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb兔单抗只识别内源性的Tyr700磷酸化c-Cbl蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr700 of human c-Cbl protein.

该单克隆抗体经合成与人c-Cbl蛋白Tyr700的邻近氨基酸残基序列一致,免疫动物产生。

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated (-) or treated with pervanadate (1 mM; +), using Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (upper) or c-Cbl (D4E10) Rabbit mAb #8447 (lower). Western blot检测经pervanadate (1 mM; +)处理或未经pervanadate处理的Jurkat细胞。采用Phospho-c-Cbl (Tyr700) (D16D7)Rabbit mAb (upper) 或c-Cbl (D4E10) Rabbit mAb #8447 (lower)。

Specificity

Specificity

Western blot analysis of extracts from Jurkat cells, untreated (-) or treated with pervanadate (1 mM; +), using Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (left). The phospho-specificity of the antibody was verified by preincubating the antibody with c-Cbl (Tyr700) phosphopeptide (center) or with c-Cbl (Tyr700) nonphosphopeptide prior to incubation with the membrane (right). Western blot检测经pervanadate (1 mM; +)处理或未经pervanadate处理的Jurkat细胞,采用Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (left)。在与细胞膜作用前,抗体预先与c-Cbl (Tyr700)磷酸肽(center)或c-Cbl (Tyr700)非磷酸肽孵育,表明抗体具有磷酸特异性。

IP

IP

Immunoprecipitation (IP) of phosphorylated c-Cbl from Jurkat cell extracts, untreated (-) or treated with pervanadate (1 mM; +), using Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (lanes 4 and 6). Western blot detection was performed using the same antibody. Lanes 1 and 2 are 10% input. Lanes 3 and 5 are IPs performed with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. 免疫共沉淀检测Jurkat细胞提取物中经过pervanadate (1 mM; +)处理或未经pervanadate处理的磷酸化的c-Cbl,采用Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (lanes 4 and 6)。Western blot检测采用相同的抗体。Lanes 1和2样品浓度为10%。Lanes 3和5 为免疫共沉淀采用Rabbit (DA1E) mAb IgG XP® 同型对照#3900。

Background

The c-Cbl proto-oncogene is a ubiquitously expressed cytoplasmic adaptor protein that is especially predominant in hematopoietic cells (1,2). c-Cbl is rapidly tyrosine-phosphorylated in response to stimulation of a variety of cell-surface receptors and becomes associated with a number of intracellular signaling molecules such as protein tyrosine kinases, phosphatidylinositol-3 kinase, Crk, and 14-3-3 proteins (3,4). c-Cbl possesses a highly conserved amino-terminal phosphotyrosine binding domain (TKB) and a C3HC4 RING finger motif. The TKB recognizes phosphorylated tyrosines on activated receptor tyrosine kinases (RTKs) as well as other nonreceptor tyrosine kinases. The RING finger motif recruits ubiquitin-conjugating enzymes. These two domains are primarily responsible for the ubiquitin ligase activity of c-Cbl and downregulation of RTKs (3). Research studies have indicated that in human cancer tissues, c-Cbl is frequently tyrosine-phosphorylated in a tumor-specific manner (5). Phosphorylation of Tyr731 of c-Cbl provides a docking site for downstream signaling components such as p85 and Fyn (6).

c-Cbl原癌基因是一种广泛表达的细胞质衔接蛋白,主要表达于造血细胞(1,2)。对于许多种细胞表面受体的刺激,c-Cbl迅速地进行酪氨酸磷酸化,并与许多细胞内信号分子例如蛋白酪氨酸激酶,磷脂酰肌醇-3激酶,Crk和14-3-3蛋白相联合(3,4)。c-Cbl具有高度保守的氨基末端磷酸酪氨酸结合结构域(TKB),以及C3HC4 RING指模序。TKB识别活化的酪氨酸激酶受体(RTKs)上的磷酸化的酪氨酸,以及其他非受体酪氨酸激酶。环指模序募集泛素缀合酶。这两种结构域主要负责c-Cbl的泛素连接酶活性,以及RTKs的下调(3)。研究表明人癌症组织,c-Cbl以一种肿瘤特异的方式频繁地进行酪氨酸磷酸化(5)。c-Cbl Tyr731的磷酸化为下游的信号组分诸如p85 和 Fyn提供了一个停泊位点(6)。

It has been demonstrated that c-Cbl is phosphorylated at Tyr700 by Fyn, Yes, and Syk (4) and that Vav, a hematopoietic-restricted Rac guanine nucleotide exchange factor, undergoes c-Cbl-dependent ubiquitination upon recruitment to phospho-Tyr700 (5).

c-Cbl在Tyr700磷酸化,主要由Fyn, Yes和Syk (4)以及Vav,一种局限于造血细胞中的Rac鸟嘌呤核苷酸交换因子,募集到磷酸化的Tyr700并进行c-Cbl-依赖的泛素化作用(5)。

  1. Blake, T.J. et al. (1991) Oncogene 6, 653-657.
  2. Thien, C.B. and Langdon, W.Y. (1998) Immunol. Cell Biol. 76, 473-482.
  3. Christine, B.F. et al. (2001) Nat. Rev. Mol. Cell Biol. 2, 294-307.
  4. Feshchenko, E.A. et al. (1998) J. Biol. Chem. 273, 8323-8331.
  5. Kamei, T. et al. (2000) Int. J. Oncol. 17, 335-339.
  6. Hunter, C. et al. (1999) J. Biol. Chem. 274, 2097-2106.
  7. Miura-Shimura, Y. et al. (2003) J Biol Chem 278, 38495-504.

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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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