PhosphoPlus? Rb (Ser780, Ser807/811) Antibody Kit

• 产品编号：CST-9306S      品牌：CST       原厂货号：9306S
• 产品分类：生化和细胞分析 > 细胞分析试剂盒 > 细胞增殖及周期检测试剂盒
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描述：

Phospho-Rb (Ser780)和 Phospho-Rb (Ser807/811)能够分别识别内源性丝氨酸(780位，807/811位)磷酸化的Rb蛋白。这些抗体不与其他位点发生磷酸化的Rb蛋白发生交叉反应。Rb (4H1) Mouse mAb识别内源性Rb总蛋白。该抗体不与Rb同源蛋白p107、p130或其他蛋白发生交叉反应。多克隆抗体是由合成的人源的针对Rb蛋白丝氨酸(780位，807/811位)的磷酸化肽段免疫动物，采用蛋白A和多肽亲和层析技术纯化生产的。Rb (4H1) monoclonal antibody是由Rb-C Fusion Protein #6022免疫动物生产的。Rb-C包含人类Rb蛋白的701-928氨基酸残基。该PhosphoPlus® Rb (Ser780, Ser807/811) Antibody Kit提供在细胞内研究细胞周期进程的试剂和方法。该试剂盒包含一种Rb总蛋白抗体，两种不同的Rb磷酸化抗体，Rb对照蛋白，一抗和二抗的量足够10次western blot试验。 视网膜母细胞瘤肿瘤抑制蛋白Rb通过控制细胞周期的G1期限制点的进程来调控细胞增殖(1)。Rb具有3个功能特异的结合结构域并能够与转录因子家族的关键调节蛋白E2F、c-Abl酪氨酸激酶和具有保守的LXCXE模体蛋白相互作用(2-4)。CDK介导的细胞周期依赖性磷酸化能够抑制Rb靶向结合并允许细胞周期进程(5)。Rb的失活和随后的细胞周期进程似乎需要伴随周期蛋白E-CDK2磷酸化的cyclin D-CDK4/6初始磷酸化(6)。离体已经观察到不同CDK/cyclin复合物的特异性(6-8)，而且周期蛋白D1为第780位丝氨酸在体磷酸化所必需(9)。

1. Sherr, C.J. (1996) Science 274, 1672-7.
2. Nevins, J.R. (1992) Science 258, 424-9.
3. Welch, P.J. and Wang, J.Y. (1993) Cell 75, 779-90.
4. Hu, Q.J. et al. (1990) EMBO J 9, 1147-55.
5. Knudsen, E.S. and Wang, J.Y. (1997) Mol Cell Biol 17, 5771-83.
6. Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61.
7. Connell-Crowley, L. et al. (1997) Mol Biol Cell 8, 287-301.
8. Kitagawa, M. et al. (1996) EMBO J 15, 7060-9.
9. Geng, Y. et al. (2001) Proc Natl Acad Sci USA 98, 194-9.

原厂资料：

Specificity / Sensitivity

Phospho-Rb (Ser780) and Phospho-Rb (Ser807/811) antibodies detect endogenous levels of Rb only when phosphorylated at Ser780 and Ser807/811, respectively. These antibodies do not cross-react with Rb phosphorylated at other sites. Rb (4H1) Mouse mAb detects endogenous levels of total Rb protein. The antibody does not cross-react with the Rb homologues p107 or p130, or with other proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser780 and Ser807/811 of human Rb. Antibodies are purified by protein A and peptide affinity chromatography. Rb (4H1) monoclonal antibody is produced by immunizing animals with Rb-C Fusion Protein #6022, which contains residues 701-928 of human Rb .

Description

The PhosphoPlus® Rb (Ser780, Ser807/811) Antibody Kit provides reagents and protocols to investigate cell cycle progression within cells. The kit contains a total Rb antibody, two distinct phospho-Rb antibodies, and Rb control proteins along with secondary antibodies and reagents to perform up to 10 western blots.

Background

The retinoblastoma tumor suppressor protein, Rb, regulates cell proliferation by controlling progression through the restriction point within the G1-phase of the cell cycle (1). Rb has three functionally distinct binding domains and interacts with critical regulatory proteins including the E2F family of transcription factors, c-Abl tyrosine kinase and proteins with a conserved LXCXE motif (2-4). Cell cycle-dependent phosphorylation by a CDK inhibits Rb target binding and allows cell cycle progression (5). Rb inactivation and subsequent cell cycle progression likely requires an initial phosphorylation by cyclin D-CDK4/6 followed by cyclin E-CDK2 phosphorylation (6). Specificity of different CDK/cyclin complexes has been observed in vitro (6-8) and cyclin D1 is required for Ser780 phosphorylation in vivo (9). 

1. Sherr, C.J. (1996) Science 274, 1672-7.
2. Nevins, J.R. (1992) Science 258, 424-9.
3. Welch, P.J. and Wang, J.Y. (1993) Cell 75, 779-90.
4. Hu, Q.J. et al. (1990) EMBO J 9, 1147-55.
5. Knudsen, E.S. and Wang, J.Y. (1997) Mol Cell Biol 17, 5771-83.
6. Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61.
7. Connell-Crowley, L. et al. (1997) Mol Biol Cell 8, 287-301.
8. Kitagawa, M. et al. (1996) EMBO J 15, 7060-9.
9. Geng, Y. et al. (2001) Proc Natl Acad Sci USA 98, 194-9.

注意事项：

Antibody 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 antibodies.