Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb #8516

No. Size Price
8516S 100 µl ( 10 western blots ) ¥4,050.00 现货查询 购买询价
8516T 20 µl ( 2 western blots ) ¥1,600.00 现货查询 购买询价
8516 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 110 Rabbit IgG
IP 1:200
IHC-P 1:400
F 1:400
IF-IC 1:1600

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),

Specificity / Sensitivity

Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb recognizes endogenous levels of Rb protein only when phosphorylated at Ser807, Ser811, or at both sites. This antibody does not cross-react with Rb phosphorylated at Ser608.Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb能够识别内源性丝氨酸(807位),丝氨酸(811位)或两个位点同时磷酸化的Rb蛋白。该抗体不与丝氨酸(608)磷酸化的Rb蛋白发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser807/811 of human Rb protein.该单克隆抗体是由合成的人源的针对Rb蛋白丝氨酸(807/811位)的磷酸化肽段免疫动物生产的。

IF-IC

IF-IC

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 (Alexa Fluor® 488 Conjugate) (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb.免疫组织化学方法检测石蜡包埋人类结肠癌,使用的抗体为Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb 。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human ovarian serous adenocarcioma using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).免疫组织化学方法检测石蜡包埋人类卵巢浆液性腺癌,使用的抗体为Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb ,左图为共孵育对照多肽,右图为共孵育抗原特异性多肽。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb.免疫组织化学方法检测石蜡包埋人类肺癌,左图为对照组,右图为λ磷酸酶处理组,使用的抗体为Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb。

Western Blotting

Western Blotting

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 方法检测MCF7细胞提取物,分为未处理组(-)或小牛肠碱性磷酸酶及λ磷酸酶处理组 (+), 使用的抗体为Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb (上图) 或Rb (4H1) Mouse mAb #9309 (下图).

Western Blotting

Western Blotting

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.Western blot 方法检测WI-38细胞提取物,分为血清饥饿3天组(-)或血清饥饿3天后10%血清处理2天组(+),使用的抗体为Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb。

IP

IP

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.使用Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb 从MCF7细胞提取物中免疫沉淀丝氨酸(807/811位)的磷酸化Rb蛋白(泳道2)。Western blot使用同一抗体。泳道1为10%input。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb compared to Propidium Iodide (PI)/RNase Staining Solution #4087.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse spleen using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb.

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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