| 货号 | 12349S |
| 反应种属 | Human/Mouse/Rat/Monkey/Zebrafish |
| 来源宿主 | Rabbit |
| 应用 | W/IF-IC/ChIP |
| 目标/特异性 | Histone H2A (D6O3A) Rabbit mAb recognizes endogenous levels of total histone H2A protein. This antibody does not cross-react with other histone proteins. |
| 使用方法 | WB(1:1000) IF-IC (1:200) ChIP (1:100) |
| 供应商 | CST |
| 背景 | Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11). |
| 存放说明 | -20C |
| 计算分子量 | 14 |
Confocal immunofluorescent analysis of HeLa cells using Histone H2A (D6O3A) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).采用共聚焦免疫荧光术检测Hela细胞，使用的抗体为Histone H2A (D6O3A) Rabbit mAb (绿色)。肌动蛋白微丝使用DY-554 phalloidin进行标记（红色）。 | |
Western blot analysis of extracts from various cell lines using Histone H2A (D6O3A) Rabbit mAb.Western blot方法检测多种细胞系的提取物，使用的抗体为Histone H2A (D6O3A) Rabbit mAb。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 5 μl of Histone H2A (D6O3A) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MYT-1 Exon 1 Primers #4493, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.使用从4 x 106 MCF7细胞中提取的交联过的染色质，以及5 µl Histone H2A (D6O3A) Rabbit mAb 或2 µl Normal Rabbit IgG #2729进行染色质免疫沉。所用试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。富集到的DNA经过实时PCR定量，所使用引物为SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MYT-1 Exon 1 Primers #4493, 以及SimpleChIP® Human α Satellite Repeat Primers #4486。每个样品中免疫沉淀得到的DNA量由与input chromatin( 相当于1)的相对信号值来表示。 |
