| 货号 | 2187S |
| 反应种属 | Human |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC |
| 目标/特异性 | Phospho-CENP-A (Ser7) Antibody detects endogenous levels of human CENP-A protein only when phosphorylated on Ser7. This antibody does not cross-react with other histone proteins, including Histone H3. |
| 使用方法 | WB(1:1000) IP (1:25) IF-IC (1:100) |
| 供应商 | CST |
| 背景 | Modulation of chromatin structure plays a critical role in the regulation of transcription and replication of the eukaryotic genome. The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. In addition to the growing number of post-translational histone modifications regulating chromatin structure, cells can also exchange canonical histones with variant histones that can directly or indirectly modulate chromatin structure (1). CENP-A, also known as the chromatin-associated protein CSE4 (capping-enzyme suppressor 4-p), is an essential histone H3 variant that replaces canonical histone H3 in centromeric heterochromatin (2). The greatest divergence between CENP-A and canonical histone H3 occurs in the amino-terminal tail of the protein, which binds linker DNA between nucleosomes and facilitates proper folding of centromeric heterochromatin (3). The amino-terminal tail of CENP-A is also required for recruitment of other centromeric proteins (CENP-C, hSMC1, hZW10), proper kinetochore assembly and chromosome segregation during mitosis (4). Additional sequence divergence in the histone fold domain is responsible for correct targeting of CENP-A to the centromere (5). Many of the functions of CENP-A are regulated by phosphorylation (6,7). Aurora A-dependent phosphorylation of CENP-A on Ser7 during prophase is required for proper targeting of Aurora B to the inner centromere in prometaphase, proper kinetochore/microtubule attachment and proper alignment of chromosomes during mitosis (6). |
| 存放说明 | -20C |
| 计算分子量 | 17 |
Western blot analysis of extracts from HeLa cells arrested in S phase or mitosis using Phospho-CENP-A (Ser7) Antibody (upper panel) or CENP-A Antibody #2186 (lower panel). S phase cells were treated for 12 hours with thymidine (2 mM), rinsed three times, released into normal growth medium for 10 hours and then treated an additional 12 hours with thymidine before harvesting. Mitotic cells were treated for 12 hours with thymidine, rinsed three times and then treated for 16 hours with paxitaxol (500 nM final). | |
使用Phospho-CENP-A (Ser7) Antibody (上图)或CENP-A Antibody #2186 (下图)，免疫印迹(Western blot)分析停滞的S期或有丝分裂的HeLa细胞中Phospho-CENP-A (Ser7)和CENP-A蛋白水平。S期的细胞经过thymidine (2 mM)处理12小时，冲洗三次后，使用普通的细胞培养液培养10小时，然后在收集细胞之前用thymidine处理12小时。有丝分裂期细胞经过thymidine处理12小时，冲洗三次后，然后用paxitaxol处理 (最终浓度为500 nM )16小时。 |
