Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Rb (4H1) Mouse mAb #9309

Rb 4H1   retinoblastoma   sc-50  

No. Size Price
9309L 300 µl ( 60 western blots ) ¥7,504.00 现货查询 购买询价
9309S 100 µl ( 20 western blots ) ¥3,100.00 现货查询 购买询价
9309T 20 µl ( 2 western blots ) ¥1,200.00 现货查询 购买询价
9309 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:2000 Human,Monkey,Bovine,Pig, Endogenous 110 Mouse IgG2a
IP 1:100
IHC-P 1:100
F 1:400
IF-IC 1:800
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Specificity / Sensitivity

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.Rb (4H1) Mouse mAb 能够检测内源性Rb总蛋白。该抗体不与Rb同源蛋白p107或p130或其他蛋白发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with Rb-C Fusion Protein #6022, containing residues 701-928 of human Rb .该单克隆抗体是由Rb-C Fusion Protein #6022包含人类Rb蛋白701-928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 COS-7 cells, untreated or hydroxyurea-treated (G1/S), using Rb (4H1) Mouse mAb.Western blot方法检测COS-7细胞提取物,分为非处理组或羟基脲处理(G1/S)组,使用的抗体为 Rb (4H1) Mouse mAb。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Rb (4H1) Mouse mAb.免疫组织化学方法检测石蜡包埋人类乳腺癌组织,示核定位,使用的抗体为Rb (4H1) Mouse mAb。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Rb (4H1) Mouse mAb.免疫组织化学方法检测石蜡包埋人类肺癌组织,使用的抗体为Rb (4H1) Mouse mAb。

IF-IC

IF-IC

Confocal immunofluorescent image of SH-SY5Y cells, using RB (4H1) Mouse mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).激光共聚焦免疫荧光方法检测SH-SY5Y细胞,使用的抗体为RB (4H1) Mouse mAb ,呈绿色。肌动蛋白纤维由Alexa Fluor® 555 鬼笔环肽标记,为红色。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, using Rb (4H1) Mouse mAb versus propidium iodide (DNA content). The box indicates Rb positive cells.流式细胞术检测Jurkat细胞,使用的抗体为Rb (4H1) Mouse mAb ,碘化乙啶检测DNA含量作为对照。框内示Rb阳性细胞。

Western Blotting

Western Blotting

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方法分析Hela细胞提取物。分别用100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-), SignalSilence® Rb siRNA I #6451 (+)或SignalSilence® Rb siRNA II (+)转染HeLa细胞。使用的抗体为Rb (4H1) Mouse mAb #9309和α-Tubulin (11H10) Rabbit mAb #2125。和对照抗体α-Tubulin (11H10) Rabbit mAb #2125相比,Rb (4H1) Mouse mAb的检测结果证实Rb的表达被Rb siRNA有效沉默。

Chromatin IP

Chromatin IP

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.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 (4H1) Mouse 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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