| 货号 | 9969T |
| 描述 | The Rb Antibody Sampler Kit provides reagents and protocols to investigate cell cycle progression within cells. The kit contains primary and secondary antibodies to perform four Western blot experiments with each antibody.该Rb Antibody Sampler Kit 提供在细胞内研究细胞周期的试剂和方法。该试剂盒包含足够四次western blot的一抗和二抗。 |
| 目标/特异性 | All antibodies contained in this kit detect endogenous levels of their respective target protein. |
| 供应商 | CST |
| 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 |
| 参考文献 | Sherr, C.J. (1996) Science 274, 1672-7. Nevins, J.R. (1992) Science 258, 424-9. Welch, P.J. and Wang, J.Y. (1993) Cell 75, 779-90. Hu, Q.J. et al. (1990) EMBO J 9, 1147-55. Knudsen, E.S. and Wang, J.Y. (1997) Mol Cell Biol 17, 5771-83. Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61. Connell-Crowley, L. et al. (1997) Mol Biol Cell 8, 287-301. Kitagawa, M. et al. (1996) EMBO J 15, 7060-9. Geng, Y. et al. (2001) Proc Natl Acad Sci USA 98, 194-9. |
Immunohistochemical analysis of paraffin-embedded mouse spleen using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb. | |
Western blot analysis of extracts from WI-38 cells, serum-starved for 3 days (-) or serum-starved for 3 days followed by treatment with 10% serum for 2 days (+), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb. | |
Immunoprecipitation of phospho-Rb (Ser807/811) from MCF7 cell extracts using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb (lane 2). Western blot was performed using the same antibody. Lane 1 is 10% input. | |
Flow cytometric analysis of Jurkat cells using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb compared to Propidium Iodide (PI)/RNase Staining Solution #4087. | |
Western blot analysis of extracts from MCF7 cells, untreated (-) or treated with calf intestinal phosphatase (CIP) and λ phosphatase (+), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb (upper) or Rb (4H1) Mouse mAb #9309 (lower). | |
Western blot analysis of phosphorylated or nonphosphorylated recombinant, truncated Rb, without or with Rb blocking peptide, using Phospho-Rb (Ser780) (D59B7) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb. | |
Western blot analysis of extracts from WI-38 cells, serum-starved for 3 days (-) or serum-starved for 3 days followed by treatment with 10% serum for 2 days (+), using Phospho-Rb (Ser780) (D59B7) Rabbit mAb. | |
Immunoprecipitation of phospho-Rb (Ser780) from WI-38 cell extracts using Phospho-Rb (Ser780) (D59B7) Rabbit mAb (lane 2). Western blot was performed using the same antibody. Lane 1 is 10% input. | |
Immunohistochemical analysis of paraffin-embedded human ovarian serous adenocarcinoma using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right). | |
Confocal immunofluorescent analysis of MCF7 (left) and BT-549 (right) cells, untreated (upper) or λ phosphatase-treated (lower) using Phospho-Rb (Ser807/Ser811) (D20B12) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Raji cells and either 5 μl of Rb (4H1) Mouse 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. | |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-), SignalSilence® Rb siRNA I #6451 (+) or SignalSilence® Rb siRNA II (+), using Rb (4H1) Mouse mAb #9309 and α-Tubulin (11H10) Rabbit mAb #2125. The Rb (4H1) Mouse mAb confirms silencing of Rb expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Rb siRNA. | |
Western blot analysis of Rb Control Protein #9303 using Phospho-Rb (Ser795) Antibody #9301 (upper) or Rb (4H1) mAb #9309 (lower).Western blot 方法检测Rb Control Protein #9303 ，使用的抗体为Phospho-Rb (Ser795) Antibody #9301 (上图) 或 Rb (4H1) mAb #9309 (下图)。 |
