Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Rb (D20) Rabbit mAb #9313

rb   sc-50  

No. Size Price
9313S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
9313T 20 µl ( 2 western blots ) ¥1,200.00 现货查询 购买询价
9313 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Monkey, Endogenous 110 Rabbit
IP 1:100
ChIP 1:200

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP,

Specificity / Sensitivity

Rb (D20) Rabbit mAb detects endogenous levels of total Rb protein.Rb (D20) Rabbit mAb 能够检测内源性Rb总蛋白、

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to central residues of human Rb.该单克隆抗体是由合成的人源的针对Rb蛋白中间氨基酸残基的肽段免疫动物生产的。

Western Blotting

Western Blotting

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。

IP

IP

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。

Chromatin IP

Chromatin IP

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

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). 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.

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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