| 货号 | 9309L |
| 反应种属 | Human/Monkey/Bovine/Pig |
| 来源宿主 | Mouse |
| 应用 | W/IP/IHC-P/IF-IC/F/ChIP |
| 使用方法 | WB(1:2000) IP (1:100) IHC-P (1:100) F (1:400) IF-IC (1:800) ChIP (1:100) |
| 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 |
| 计算分子量 | 110 |
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。 | |
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。 | |
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。 | |
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 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 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 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。 |
