| 货号 | 4353S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/ChIP |
| 目标/特异性 | Acetyl-Histone H3 (Lys27) Antibody detects endogenous levels of histone H3 when acetylated on Lys27. This antibody shows weak cross-reactivity with histone H3 acetylated on Lys9. This antibody does not cross-react with Histone H3 acetylated on lysines 14, 18 and 56. |
| 使用方法 | WB(1:1000) IP (1:25) ChIP (1:25) |
| 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 | 17 |
Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or treated with Trichostatin A #9950 (400 nM for 18 h), using Acetyl-Histone H3 (Lys27) Antibody (upper) and Histone H3 Antibody #9715 (lower). 使用Acetyl-Histone H3 (Lys27) Antibody (上图)和Histone H3 Antibody #9715 (下图)，免疫印迹(Western blot)分析HeLa和NIH/3T3细胞中Acetyl-Histone H3 (Lys27)和Histone H3蛋白水平，细胞分为untreated或Trichostatin A #9950 (400 nM for 18 h)。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 20 μl of Acetyl-Histone H3 (Lys27) Antibody 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 AFM Intron 1 Primers #5098, 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SimpleChIP®Enzymatic Chromatin IP Kit (Magnetic Beads) #9003，用4 x 106 HeLa细胞的交联染色质以及20 µl Acetyl-Histone H3 (Lys27) Antibody或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP® Human GAPDH Exon 1 Primers #5516、SimpleChIP® Human RPL30 Exon 3 Primers #7014、SimpleChIP® Human AFM Intron 1 Primers #5098和SimpleChIP® Human α Satellite Repeat Primers #4486，浓缩的DNA通过real-time PCR定量。在每个样品中免疫沉淀DNA的数量被当做一个相对于总input chromatin的数量的信号，这相当于一。 |
