| 货号 | 8056S |
| 反应种属 | Human/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/ChIP |
| 目标/特异性 | SIN3A (D9D6) Rabbit mAb recognizes endogenous levels of total SIN3A protein. Based on protein sequence, this antibody is not predicted to cross-react with SIN3B protein. |
| 使用方法 | WB(1:1000) IP (1:50) IF-IC (1:200) ChIP (1:50) |
| 供应商 | CST |
| 背景 | SIN3 was originally identified as a negative regulator of transcription in budding yeast (1,2). Since then, three isoforms of the SIN3 proteins have been identified in mammalian cells, as products of two different genes, SIN3A and SIN3B (3,4). Both SIN3A and SIN3B are nuclear proteins that function as scaffolding subunits for the multi-subunit SIN3 transcriptional repressor complex, containing SIN3A or SIN3B, HDAC1, HDAC2, SDS3, RBBP4/RBAP48, RBBP7/RBAP46, SAP30, and SAP18 (3,4). SIN3 proteins contain four paired amphipathic alpha-helix (PAH) motifs that function in the recruitment of the SIN3 complex to target genes by binding a multitude of DNA-binding transcriptional repressor proteins, including Mad1, p53, E2F4, HCF-1, AML1, Elk-1, NRSF, CTCF, ERα, and MeCP2 (3,4). In addition, SIN3 proteins contain an HDAC interaction domain (HID), which mediates binding of HDAC1 and HDAC2 via the SDS3 bridging protein, and a highly conserved region (HCR) at the carboxy terminus, which contributes to repressor protein binding (3,4). RBBP4 and RBBP7 proteins also bind to SDS3 and contribute to nucleosome binding of the complex. The SIN3 complex functions to repress transcription, in part, by deacetylating histones at target gene promoters (3,4). In addition, recent studies have shown that SIN3 is recruited to the coding regions of repressed and active genes, where it deacetylates histones and suppresses spurious transcription by RNA polymerase II (3,5). In addition to histone deacetylase activity, the SIN3 complex associates with histone methyltransferase (ESET), histone demethylase (JARID1A/RBP2), ATP-dependent chromatin remodeling (SWI/SNF), methylcytosine dioxygenase (TET1), and O-GlcNAc transferase (OGT) activities, all of which appear to contribute to the regulation of target genes (5-9). The SIN3 complex is critical for proper regulation of embryonic development, cell growth and proliferation, apoptosis, DNA replication, DNA repair, and DNA methylation (imprinting and X-chromosome inactivation) (3,4). |
| 存放说明 | -20C |
| 计算分子量 | 145 |
Confocal immunofluorescent analysis of 293 cells using SIN3A (D9D6) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). 使用SIN3A (D9D6) Rabbit mAb兔单抗 (绿色)，共聚焦免疫荧光分析293细胞。DY-554 phalloidin标记微丝蛋白(红色)。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells grown in phenol red free medium and charcoal stripped FBS for 4 days then treated with β-estradiol (10 nM) for 1 hr and either 10 μl of SIN3A (D9D6) 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 ESR1 Promoter Primers #9673, SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD Exon 1 Primers #4490, 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，来自在无酚红培养基和葡聚糖处理胎牛血清中培养4天之后处理了β-estradiol (10 nM) 1小时的4 x 106 MCF7细胞的交联染色质以及10µl SIN3A (D9D6) Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP® Human ESR1 Promoter Primers #9673、SimpleChIP® Human GAPDH Exon 1 Primers #5516、SimpleChIP® Human MyoD Exon 1 Primers #4490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。 | |
Western blot analysis of extracts from various cell lines using SIN3A (D9D6) Rabbit mAb. 使用SIN3A (D9D6) Rabbit mAb兔单抗，免疫印迹(Western blot)分析不同细胞中SIN3A蛋白水平。 |
