| 货号 | 9932T |
| 描述 | Cell Cycle Regulation Antibody Sampler kit offers an economical way of detecting eight integral cell cycle regulation proteins. The kit contains enough primary and secondary antibodies to perform four Western blot experiments with each primary antibody.Cell Cycle Regulation Antibody Sampler kit为研究8种必须的细胞周期调节蛋白提供了一个经济的方式。该试剂盒提供了足够4次western blot实验的一抗和二抗。 |
| 目标/特异性 | Antibodies detect endogenous levels of their respective proteins. |
| 供应商 | CST |
| 背景 | Eukaryotic cell cycle progression is dependent, in part, on the tightly regulated activity of cyclin dependent kinases (CDKs). Cyclin D/CDK4/6 activity occurs in mid-late G1 phase, upstream of CDK2/cyclin E activity. Both of these activities are required for hyperphosphorylation of the retinoblastoma gene product (pRb). pRb phosphorylation allows the release of S phase-promoting transcription factors and is indicative of the cells commitment to proliferate. This point in the cell cycle is known as the restriction point. Cyclin protein levels oscillate throughout the cell cycle, and their availability is a means of controlling CDK activity and cell proliferation. Cyclin D is degraded through the ubiquitin proteasome pathway in the absence of mitogenic signaling. Ubiquitination of cyclin D1 is enhanced by phosphorylation at Thr286 by glycogen synthase kinase 3b (GSK-3b) (1). p27/Kip1, p57 Kip2 and p21 Waf1/Cip1 are members of the Cip/Kip family of cyclin-dependent kinase inhibitors. They form heterotrimeric complexes with cyclins and CDKs, inhibiting kinase activity and blocking progression through G1/S phase (2). However, p21 may enhance assembly and activity of cyclin D/CDK4/6 complexes (3). Levels of p21 and p27 protein are controlled through ubiquitination and proteasomal degradation (4). Levels of p27 are upregulated in quiescent cells and in cells treated with negative cell cycle regulators. p27 nuclear localization is controlled by Akt-dependent phosphorylation at Thr157 (5). The inhibitors of CDK4 (INK4) family include p15 INK4B, p16 INK4A, p18 INK4C, and p19 INK4D. All INK4 proteins selectively inhibit CDK4/6 activity, either in a binary complex, or in a ternary complex including cyclin D, resulting in inhibition of cell division (6,7).真核细胞的细胞周期进程部分依赖细胞周期蛋白依赖性激酶(CDKs)的紧密调节活性。Cyclin D/CDK4/6的活性出现在G1中后期和CDK2/cyclin E活性的上游。这些活性都需要视网膜母细胞瘤基因产物(pRb)的过度磷酸化。pRb磷酸化可以释放S期促进转录因子和表明细胞增殖。细胞周期中的这一点被称作限制点。细胞周期蛋白水平在整个细胞周期振荡，其有效性是控制CDK活性和细胞增殖的一种方式。有丝分裂信号通路中细胞周期蛋白D通过泛素蛋白酶体途径降解。原合酶激酶3b (GSK-3b)磷酸化Thr286能够增强细胞周期蛋白D1的泛素化(1)。p27/Kip1，p57 Kip2 和p21 Waf1/Cip1p27/Kip1是细胞周期蛋白依赖性激酶抑制剂Cip/Kip家族的成员。它们与周期蛋白和CDKs形成异源三聚体复合物，抑制细胞周期蛋白激酶活性和阻止通过G1/S期的进程(2)。然而，p21可以提高细胞周期蛋白D/CDK4/6复合物装配和活性(3)。p21和p27蛋白的水平通过泛素化和蛋白酶体降解来控制(4)。静止期细胞和阴性细胞周期调节子中p27蛋白水平上调。Akt依赖的Thr157磷酸化控制p27蛋白核定位(5)。CDK4抑制剂(INK4)家族包括p15 INK4B，p16 INK4A，p18 INK4C和p19 INK4D。所有INK4蛋白选择性抑制CDK4/ 6的活性，无论是在二元复合物中，还是在包括细胞周期蛋白D的三元复合物中，都能够抑制细胞分裂(6,7)。 |
| 存放说明 | -20C |
| 参考文献 | Diehl, J.A. et al. (1997) Genes Dev. 11, 957-972. Pestell, R.G. et al. (1999) Endocrine Rev. 20, 501-534. Cheng, J. et al. (1999) EMBO J. 18, 1571-1573. Sheaff, R.J. et al. (2000) Cell 5, 403-410. Shin, I. et al. (2002) Nat. Med. 8, 1145-1152. Guan, K.L. et al. (1994) Genes Dev. 8, 2939-2952. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681. |
Flow cytometric analysis of Jurkat cells using CDK4 (D9G3E) Rabbit mAb and Propidium Iodid (PI)/RNase Staining Solution #4087 to measure DNA content. Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using CDK4 (D9G3E) Rabbit mAb. | |
Western blot analysis of extracts from various cell lines using CDK4 (D9G3E) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using CDK4 (D9G3E) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right). | |
Confocal immunofluorescent analysis of MCF7 cells using CDK4 (D9G3E) Rabbit mAb (green), p21 Waf1/Cip1 (12D1) Rabbit mAb (Alexa Fluor® 555 Conjugate) #8493 (red), and Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #3458 (blue pseudocolor). | |
Flow cytometric analysis of Jurkat cells using p27 Kip1 (D69C12) XP® Rabbit mAb versus Propidium Iodide (PI)/RNase Staining Solution #4087. Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. | |
Western blot analysis of extracts from HeLa, NIH/3T3, C6 and COS cells, using CDK2 (78B2) Rabbit mAb #2546. | |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p21 Waf1/Cip1 siRNA II (+), using p21 Waf1/Cip1 (12D1) Rabbit mAb #2947 and α-Tubulin (11H10) Rabbit mAb #2125. The p21 Waf1/Cip1 (12D1) Rabbit mAb confirms silencing of p21 Waf1/Cip1 expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p21 Waf1/Cip1 siRNA. | |
Confocal immunofluorescent analysis of MCF-7 cells using p27 Kip1 (D69C12) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). | |
Immunoprecipitation of p27 Kip1 from 293 cells using p27 Kip1 (D69C12) XP® Rabbit mAb. Western analysis was performed using the same antibody. Lane 1 is 5% input. | |
Immunohistochemical analysis of paraffin-embedded HeLa cells, transfected with SignalSilence® Control siRNA (Unconjugated) #6568 (left) or SignalSilence®p21 Waf1/Cip1 siRNA II #6558 (right), using p21 Waf1/Cip1 (12D1) Rabbit mAb. | |
Western blot analysis of extracts from various cell types using p27 Kip1 (D69C12) XP® Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded Apc (min/+) mouse intestine using Cyclin D1 (92G2) Rabbit mAb. | |
Flow cytometric analysis of HeLa cells (red) and MCF7 cells (blue), using p21 Waf1/Cip1 (12D1) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using p21 Waf1/Cip1 (12D1) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right). |
