Revision 1

#9794Store at -20C

 

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Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
c-Cbl (C49H8) Rabbit mAb 2179 40 µl 120 kDa Rabbit IgG
Phospho-c-Cbl (Tyr731) Antibody 3554 40 µl 120 kDa Rabbit 
Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb 8869 40 µl 120 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The c-Cbl Antibody Sampler Kit provides an economical means of evaluating levels of c-Cbl protein phosphorylated at the specified sites, as well as total c-Cbl levels. The kit contains enough reagents to perform four western blot experiments per primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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).

  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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    限制使用

    除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

    专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

    Revision 1
    #9794

    c-Cbl Antibody Sampler Kit

    c-Cbl Antibody Sampler Kit: Image 1 Expand Image
    使用 c-Cbl (C49H8) Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹分析。
    c-Cbl Antibody Sampler Kit: Image 2 Expand Image
    使用 Phospho-c-Cbl (Tyr731) Antibody,对未经处理或经小牛肠磷酸酶 (CIP) 处理以及使用 c-Cbl 与 Fyn (泳道3 与6) 或不使用 Fyn (泳道2 与 5) 转染的 293-T 细胞提取物进行蛋白质印迹分析。(裂解物的提供者为马萨诸塞州哈佛大学 Dr. Hamid Band 实验室的 A.L. Reddi 博士)
    c-Cbl Antibody Sampler Kit: Image 3 Expand Image
    一抗与靶标蛋白结合之后,与偶联 HRP 的二抗形成复合体。添加 LumiGLO®,在酶催化分解期间发光。
    c-Cbl Antibody Sampler Kit: Image 4 Expand Image
    使用 Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (上图) 或 c-Cbl (D4E10) Rabbit mAb #8447(下图),对未经处理 (-) 或用钒酸钠 (1mM; +) 处理的 Jurkat 细胞提取物进行蛋白质印迹分析。
    c-Cbl Antibody Sampler Kit: Image 5 Expand Image
    使用 Phospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (左图),对未经处理 (-) 或用钒酸钠 (1mM; +) 处理的 Jurkat 细胞提取物进行蛋白质印迹分析。在该抗体孵育前 (右图),使用 c-Cbl (Tyr700) 磷酸肽 (中图) 或 c-Cbl (Tyr700) 非磷酸肽对抗体进行预孵育,来验证抗体的磷酸化特异性。
    c-Cbl Antibody Sampler Kit: Image 6 Expand Image
    使用 hospho-c-Cbl (Tyr700) (D16D7) Rabbit mAb (泳道4和6),对未经处理 (-) 或用钒酸钠 (1mM; +) 处理的 Jurkat 细胞提取物的磷酸化 c-Cbl 进行免疫沉淀 (IP)。使用相同的抗体进行蛋白质印迹检测。泳道 1 和 2 为 10% input。泳道3和5是使用兔 (DA1E) mAb IgG XP ®同种型对照#3900进行的 IP。