货号 | 3300T |
同种亚型 | Rabbit IgG |
反应种属 | Human,Monkey, |
来源宿主 | Rabbit IgG |
应用 | WB, IP , F , IF-IC |
目标/特异性 | Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb detects endogenous levels of cyclin D1 only when phosphorylated at Thr286. The antibody does not cross-react with other cyclin D family members. |
使用方法 | WB(1:1000) IP (1:50) F (1:1600) IF-IC (1:2000) |
供应商 | CST |
灵敏度 | Endogenous |
背景 | Activity of the cyclin-dependent kinases CDK4 and CDK6 is regulated by T-loop phosphorylation, by the abundance of their cyclin partners (the D-type cyclins), and by association with CDK inhibitors of the Cip/Kip or INK family of proteins (1). The inactive ternary complex of cyclin D/CDK4 and p27 Kip1 requires extracellular mitogenic stimuli for the release and degradation of p27 concomitant with a rise in cyclin D levels to effect progression through the restriction point and pRb-dependent entry into S-phase (2). The active complex of cyclin D/CDK4 targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (3). Levels of cyclin D protein drop upon withdrawal of growth factors through downregulation of its protein expression and through phosphorylation-dependent degradation (4).细胞周期蛋白依赖性蛋白激酶CDK4和CDK6的活性受三个因素的调控:T-环路磷酸化,周期蛋白伴侣(D-型周期蛋白)的丰度,与CDK抑制剂Cip/Kip 或 INK蛋白家族的关系(1)。失活的三元复合物cyclin D/CDK4 和p27 Kip1需要胞外有丝分裂刺激因子参与p27伴侣的释放和降解,伴随周期蛋白D的水平升高,通过限制点和pRb依赖性的S期进入影响细胞周期的进程(2)。活跃的cyclin D/CDK4复合体靶向视网膜母细胞瘤蛋白以利于磷酸化,促进可以激活G1/S期基因表达的转录因子E2F的释放(3)。生长因子退出后,通过下调蛋白表达和磷酸化依赖性蛋白降解,周期蛋白D的水平下降(4)。 |
存放说明 | -20C |
计算分子量 | 36 |
参考文献 | 1 . Hirai, H. et al. (1995) Mol Cell Biol 15, 2672-81. 2 . Sherr, C.J. (1996) Science 274, 1672-7. 3 . Lukas, J. et al. (1996) Mol Cell Biol 16, 6917-25. 4 . Diehl, J.A. et al. (1997) Genes Dev 11, 957-72. 5 . Sicinski, P. et al. (1995) Cell 82, 621-630. 6 . Shao, J. et al. (2000) J Biol Chem 275, 22916-24. |
Confocal immmunofluorescent analysis of HT-1080 (top) and Saos-2 cells (bottom), untreated (left) or treated with MG132 alone (center) or with MG132 followed by λ-phosphatase (right), using Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). 激光共聚焦荧光法检测HT-1080 (上) and Saos-2 cells (下), 左图为对照细胞,中间图为MG132单独处理,右图为MG132处理后加入λ磷酸酶,检测抗体为Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb,呈绿色。肌动蛋白丝以DY-554 phalloidin标记,为红色。 | |
Flow cytometric analysis of HT-1080 cells, untreated (blue) or MG132-treated (green), using Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb.流式细胞术分析未处理(蓝色)或MG-132处理的(绿色)HT-1080细胞,使用的抗体为Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb。 | |
Western blot analysis of extracts from HT-1080 cells, treated with MG132 and with or without λ phosphatase, using Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb (upper) or Cyclin D1 (DC56) Mouse mAb #2926 (lower). western blot方法检测加入或未加入λ磷酸酶的情况下,经MG132处理的HT-1080细胞提取物,使用的抗体为Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb(上图)或Cyclin D1 (DC56) Mouse mAb #2926 (下图)。 | |
Western blot analysis of extracts from HT-1080 cells, untreated or treated with MG132 (10 μM, 4 hours), using Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb. western blot方法分析未处理或MG132处理的 (10 μM, 4 hours)HT-1080细胞提取物,使用的抗体为Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb。 |