| 货号 | 13495S |
| 反应种属 | Human |
| 来源宿主 | Rabbit |
| 应用 | W |
| 使用方法 | WB(1:1000) |
| 供应商 | CST |
| 背景 | Cyclins are a family of proteins that activate specific cyclin-dependent kinases required for progression through the cell cycle. The entry of all eukaryotic cells into mitosis is regulated by activation of cdc2/cdk1 at the G2/M transition. This activation is a multi-step process that begins with the binding of the regulatory subunit, cyclin B1, to cdc2/cdk1 to form the mitosis-promoting factor (MPF). MPF remains in the inactive state until phosphorylation of cdc2/cdk1 at Thr161 by cdk activating kinase (CAK) (1,2) and dephosphorylation of cdc2/cdk1 at Thr14/Tyr15 by cdc25C (3-5). Five cyclin B1 phosphorylation sites (Ser116, 126, 128, 133, and 147) are located in the cytoplasmic retention signal (CRS) domain and are thought to regulate the translocation of cyclin B1 to the nucleus at the G2/M checkpoint, promoting nuclear accumulation and initiation of mitosis (6-9). While MPF itself can phosphorylate Ser126 and Ser128, polo-like kinase 1 (PLK1) phosphorylates cyclin B1 preferentially at Ser133 and possibly at Ser147 (6,10). At the end of mitosis, cyclin B1 is targeted for degradation by the anaphase-promoting complex (APC), allowing for cell cycle progression (11). Research studies have shown that cyclin B1 is overexpressed in breast, prostate, and non-small cell lung cancers (12-14).细胞周期蛋白是一类激活特定细胞周期依赖性激酶以推进细胞周期的蛋白质。所有真核细胞要进入细胞周期都是G2/M转换时激活cdc2/cdk1以调控的。该激活作用分成多个步骤开始于调控亚基cdc2/cdk1与cdc2/cdk1结合以形成促有丝分裂因子（MPF）。MPF保持在失活状态直到被cdk活化激酶（CAK）通过cdc2/cdk1磷酸化161位苏氨酸（1,2），cdc25C可以对cdc2/cdk1的14/15位苏氨酸去磷酸化（3-5）。四个cyclin B1磷酸化位点（126,128,133和147位丝氨酸）位于胞质滞留信号（CRS）区域，一般认为这四个磷酸化位点可以调控cyclin B1在G2/M检验点时期的移位，促进其聚集在细胞核中并起始有丝分裂（6-9）。MPF可以自磷酸化126丝氨酸和128丝氨酸，polo-样激酶1（PLK1）选择性的磷酸细胞周期蛋白B1的133位丝氨酸以及可能磷酸化147丝氨酸(6,10).在有丝分裂的最后，cyclin B1能被后期促进复合物（APC）降解，促进细胞周期的进程（11）。研究表明cyclin B1在乳腺癌，前列腺癌和非小细胞肺癌中高表达（12-14）。 |
| 存放说明 | -20C |
| 计算分子量 | 55 |
Western blot analysis of HT-29 cell extracts, untreated (-) or treated with thymidine (2 mM, 16 hr) followed by Nocodazole #2190 (10 nM, 24 hr; +), using Phospho-Cyclin B1 (Ser116) Antibody (upper), Cyclin B1 (D5C10) XP® Rabbit mAb #12231 (middle), or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).未处理(-)或经胸苷(2 mM, 16 hr)随后经诺考达唑#2190 (10 nM, 24 hr; +)处理的HT-29细胞提取物，使用Phospho-Cyclin B1 (Ser116) Antibody (上), Cyclin B1 (D5C10) XP® Rabbit mAb #12231 (中), 或 α-Actinin (D6F6) XP® Rabbit mAb #6487 (下)进行western blot分析。 | |
Western blot analysis of HT-29 cell extracts, untreated (AS) or synchronized in S-phase by double thymidine block (2 mM) followed by release into fresh medium for the indicated time, using Phospho-Cyclin B1 (Ser116) Antibody (upper), Cyclin B1 (D5C10) XP® Rabbit mAb #12231 (middle), or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).未处理或经过双胸苷抑制(2 mM)随后经新鲜培养基处理指定时间同步在S期的HT-29细胞提取物，使用Phospho-Cyclin B1 (Ser116) Antibody (上), Cyclin B1 (D5C10) XP® Rabbit mAb #12231 (中), or α-Actinin (D6F6) XP®Rabbit mAb #6487 (下)进行western blot分析。 | |
Western blot anaylsis of HeLa cell extracts, untreated (-) or treated with Nocodazole #2190 (10 nM, 24 hr; +) using Phospho-Cyclin B1 (Ser116) Antibody (upper), Cyclin B1 (D5C10) XP® Rabbit mAb #12231 (middle), or α-Actinin (D6F6) XP® rabbit mAb #6487 (lower). Membranes were mock treated (- CIP) or Calf Intestinal Alkaline Phosphatase treated (+ CIP) after transfer.未处理(-)或诺考达唑#2190 (10 nM, 24 hr; +)处理的HeLa细胞提取物，使用Phospho-Cyclin B1 (Ser116) Antibody (上), Cyclin B1 (D5C10) XP® Rabbit mAb #12231 (中), or α-Actinin (D6F6) XP® rabbit mAb #6487 (下)进行western blot分析。转染后，细胞膜被模拟处理(- CIP)或牛小肠碱性磷酸酶处理(+ CIP)。 |
