货号 | 11881S |
反应种属 | Human/Mouse/Rat/Monkey |
来源宿主 | Rabbit |
应用 | W/IP/IF-IC |
使用方法 | WB(1:1000) IP (1:100) IF-IC (1:50) |
供应商 | CST |
背景 | The initiation of DNA replication in mammalian cells is a highly coordinated process that ensures duplication of the genome only once per cell division cycle. Origins of replication are dispersed throughout the genome and their activities are regulated via the sequential binding of pre-replication and replication factors. The origin recognition complex (ORC) is thought to bind to chromatin throughout the cell cycle (1,2). The pre-replication complex (Pre-RC) forms in late mitosis/early G1 phase with the binding of CDT1 and cdc6 to the origin, which allows binding of the heterohexameric MCM2-7 complex. The MCM complex is thought to be the replicative helicase and formation of the Pre-RC is referred to as chromatin licensing. Subsequent initiation of DNA replication requires the activation of the S-phase promoting kinases cdk2 and cdc7. Cdc7 phosphorylates MCM proteins bound to chromatin and, in conjunction with CDT1, recruits the replication factor cdc45 (3-5). Cdc45 is required for formation of pre-initiation complexes at the G1/S transition and for activation of replication origins. The level of cdc45 protein expression is associated with the proliferative status of the cell or tissue. Terminally differentiated and senescent cells lack cdc45 and highly proliferative cell lines express high levels of cdc45 (6).哺乳动物细胞的DNA复制是一个高度协调的过程以确保细胞每次分裂时基因组只复制一次。复制起始点分散在整个基因组中,它们的功能通过逐次结合的复制前和复制因子进行调控。据信起始识别复合物(ORC)在细胞周期中会一直结合在染色质上(1,2)。复制前复合物(Pre-RC)形成于有丝分裂晚期/G1早期与CDT1和cdc6结合在起始点上,并介导了MCM2-7复合物的结合。MCM复合物被认为是复制解璇酶,Pre-RC的形成则被看做是染色质许可证。随后的DNA复制起始需要S-期促进激酶cdk2和cdc7d激活。Cdc7与CDT1配合磷酸化结合在染色质上的MCM蛋白,募集复制因子cdc45(3-5)。Cdc45则需要形成G1/S转换时前-起始复合物以激活复制。cdc45蛋白表达水平和细胞或者组织的增殖状态相关。终末分化和衰老细胞缺乏cdc45而快速增殖的细胞则表达大量的cdc45(6)。 |
存放说明 | -20C |
计算分子量 | 65 |
Confocal immunofluorescent analysis of 293 (left) and WI-38 (right) cells using Cdc45 (D7G6) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).使用Cdc45 (D7G6) Rabbit mAb(绿色)和β-Actin (8H10D10) Mouse mAb #3700(红色)对293(左)和WI-38(右)细胞进行激光共聚焦免疫荧光分析。 | |
Western blot analysis of extracts from various cell lines using Cdc45 (D7G6) Rabbit mAb. 使用Cdc45 (D7G6) Rabbit mAb对多种细胞提取物进行western blot分析。 | |
Immunoprecipitation of cdc45 from Jurkat cell extracts, using Cdc45 (D7G6) Rabbit mAb (lane 2) or Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Cdc45 (D7G6) Rabbit mAb.使用Cdc45 (D7G6) Rabbit mAb (第2道)或Rabbit (DA1E) mAb IgG XP® Isotype对照 #3900 (第3道)对Jurkat细胞提取物进行免疫组化分析。第1道使用10%加入量。Western blot实验使用Cdc45 (D7G6) Rabbit mAb。 |