货号 | 9313T |
同种亚型 | Rabbit |
反应种属 | Human,Mouse,Monkey, |
来源宿主 | Rabbit |
应用 | WB, IP , ChIP |
目标/特异性 | Rb (D20) Rabbit mAb detects endogenous levels of total Rb protein. |
使用方法 | WB(1:1000) IP (1:100) ChIP (1:200) |
供应商 | CST |
灵敏度 | Endogenous |
背景 | 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).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)。 |
存放说明 | -20C |
计算分子量 | 110 |
参考文献 | 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. |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Raji cells and either 5 μl of Rb (D20) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using using SimpleChIP® Human Timeless Intron 1 Primers #7001, human DHFR promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.采用来自4 x 106 Raji细胞的交叉结合染色质进行免疫共沉淀实验,细胞通过紫外线处理(100 J/m2,恢复3小时)和5 μl Rb (D20) Rabbit mAb或者2 μl Normal Rabbit IgG #2729,使用的试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。随后通过实时PCR定量富集的DNA,使用的引物为SimpleChIP® Human Timeless Intron 1 Primers #7001, human DHFR promoter primers, 和 SimpleChIP® Human α Satellite Repeat Primers #4486.。每个样品的免疫沉淀DNA的量以相对于输入染色质总量(相当于1)的信号来表示 | |
Immunoprecipitation of Rb from Jurkat cells using Rb (D20) Rabbit mAb. Western blot was performed using the same antibody.从Jurkat细胞中用Rb (D20) Rabbit mAb免疫沉淀Rb蛋白。Western blot方法使用同一抗体。 | |
Western blot analysis of extracts from WI-38 cells, serum-starved for three days and serum-stimulated for the indicated times, using Rb (D20) Rabbit mAb.Western blot 方法检测血清饥饿3天后用血清刺激不同时间的WI-38 细胞提取物,使用的抗体为 Rb (D20) Rabbit mAb。 |