Phospho-CrkII (Tyr221) Antibody

• 产品编号：CST-3491S      品牌：CST       原厂货号：3491S
• 产品分类：抗体 > 一抗 > 磷酸化抗体
• 应用分类：

描述：

Phospho-CrkII (Tyr221) Antibody仅检测tyrosine 221位点磷酸化的内源性CrkII蛋白。该抗体与Tyr207位点磷酸化CrkL蛋白发生交叉反应，但是不能与其它tyrosine位点磷酸化蛋白质发生交叉反应。 通过人工合成人源CrkII蛋白Tyr221位点周围相应的磷酸化片段去免疫动物从而制备出多克隆抗体。通过蛋白A和多肽亲和层析纯化抗体。CrkII蛋白是一个细胞内v-Crk的同系物，该蛋白属于含有SH2-SH3-SH3结构域的受体蛋白家族，上述结构域能传递来自酪氨酸激酶的信号(1)。Crk蛋白的主要功能是通过SH2-磷酸化-酪氨酸的相互作用招募在酪氨酸激酶附近的细胞质蛋白。因此，来自Crk蛋白的输出取决于SH3结合蛋白，这包括C3G和Sos鸟嘌呤核苷酸交换蛋白、Abl酪氨酸激酶、DOCK180和一些 STE20相关激酶。Crk结合蛋白的种类表明Crk蛋白的多种功能(2)。CrkII蛋白两个 SH3结构域通过一个54氨基酸连接区被分开，这在爪蟾属、小鸡和哺乳类CrkII蛋白中高度保守(3)。在这个区域Tyrosine 221位点通过Abl酪氨酸激酶(4)、IGF-I受体(5)和EGF受体(6)磷酸化。一旦Tyr221位点被磷酸化，CrkII蛋白在分子内折叠和SH2-pTyr 相互作用方面有一个改变，这会引起CrkII蛋白从酪氨酸激酶复合物上快速分离(3)。

1. Zvara, A. et al. (2001) Oncogene 20, 951-961.
2. Kiyokawa, E. et al. (1997) Crit. Rev. Oncog. 8, 329-342.
3. Rosen, S. K. et al. (1995) Nature 374, 477-479.
4. Amoui, M. and Miller, W.T. (2000) Cell. Signal. 12, 637-643.
5. Koval, A. P. et al. (1998) Biochem. J. 330, 923-932.
6. Hashimoto, Y. et al. (1998) J. Biol. Chem. 273, 17186-17191.

原厂资料：

Specificity / Sensitivity

Phospho-CrkII (Tyr221) Antibody detects endogenous levels of CrkII only when phosphorylated at tyrosine 221. The antibody cross-reacts with Tyr207-phosphorylated CrkL but does not cross-react with other tyrosine-phosphorylated proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr221 of human CrkII. Antibodies are purified by protein A and peptide affinity chromatography.

Background

CrkII, a cellular homologue of v-Crk, belongs to a family of adaptor proteins with an SH2-SH3-SH3 domain structure that transmits signals from tyrosine kinases (1). The primary function of Crk is to recruit cytoplasmic proteins in the vicinity of tyrosine kinases through SH2-phospho-tyrosine interaction. Thus, the output from Crk depends on the SH3-binding proteins, which include the C3G and Sos guanine nucleotide exchange proteins, Abl tyrosine kinase, DOCK180 and some STE20-related kinases. The variety of Crk-binding proteins indicates the pleiotropic function of Crk (2). The two CrkII SH3 domains are separated by a 54 amino acid linker region, which is highly conserved in Xenopus, chicken and mammalian CrkII proteins (3). Tyrosine 221 in this region is phosphorylated by the Abl tyrosine kinase (4), IGF-I receptor (5) and EGF receptor (6). Once Tyr221 is phosphorylated, CrkII undergoes a change in intramolecular folding and SH2-pTyr interaction, which causes rapid dissociation of CrkII from the tyrosine kinase complex (3).

1. Zvara, A. et al. (2001) Oncogene 20, 951-961.
2. Kiyokawa, E. et al. (1997) Crit. Rev. Oncog. 8, 329-342.
3. Rosen, S. K. et al. (1995) Nature 374, 477-479.
4. Amoui, M. and Miller, W.T. (2000) Cell. Signal. 12, 637-643.
5. Koval, A. P. et al. (1998) Biochem. J. 330, 923-932.
6. Hashimoto, Y. et al. (1998) J. Biol. Chem. 273, 17186-17191.

注意事项：

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