| 货号 | 8644S |
| 反应种属 | Human |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/ChIP |
| 使用方法 | WB(1:1000) IP (1:50) IF-IC (1:1600) ChIP (1:100) |
| 供应商 | CST |
| 背景 | Estrogen receptor α (ERα), a member of the steroid receptor superfamily, contains highly conserved DNA binding and ligand binding domains (1). Through its estrogen-independent and estrogen-dependent activation domains (AF-1 and AF-2, respectively), ERα regulates transcription by recruiting coactivator proteins and interacting with general transcriptional machinery (2).Phosphorylation at multiple sites provides an important mechanism to regulate ERα activity (3-5). Ser104, 106, 118, and 167 are located in the amino-terminal transcription activation function domain AF-1, and phosphorylation of these serine residues plays an important role in regulating ERα activity. Ser118 may be the substrate of the transcription regulatory kinase CDK7 (5). Ser167 may be phosphorylated by p90RSK and Akt (4,6). According to the research literature, phosphorylation at Ser167 may confer tamoxifen resistance in breast cancer patients (4).雌激素受体α(ERα)，是类固醇受体超家族的一员，含有高度保守的DNA结合域（DBD）和配体结合域（LBD）（1）。ERα通过其雌激素非依赖性和雌激素依赖性激活域（分别为AF-1和AF-2），调节转录招募共激活因子蛋白和与一般转录机制的相互作用（2）。磷酸化提供了调节ERα活性的重要机制（3,4）。 ERα有多个磷酸化位点（5）。丝氨酸（104，106，118和167位）都位于氨基末端的转录激活功能域AF-1，这些丝氨酸的磷酸化在调节ERα活性中起着重要作用。丝氨酸（118位）可能是转录调节激酶CDK7的底物（5）。丝氨酸（167位）可以通过p90RSK和Akt磷酸化（4,6）。 Ser167的磷酸化可能会使乳腺癌患者对他莫昔芬耐药（4）。 |
| 存放说明 | -20C |
| 计算分子量 | 66 |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 1 h and either 5 μl of Estrogen Receptor α (D8H8) 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 pS2 Promoter Primers #9702, 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.通过交联染色质进行染色质免疫沉淀实验，交联染色质来自于4 x 106个细胞，细胞在不含酚红，含有活性炭/葡聚糖处理的5%FBS中培养4天，随后用 β-雌二醇(10 nM) 处理1 h，和 5 μl Estrogen Receptor α (D8H8) Rabbit mAb或2 μl Normal Rabbit IgG #2729处理，使用的试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。通过real-time PCR 方法，对提取的DNA进行量化，使用的引物为SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702和SimpleChIP® Human α Satellite Repeat Primers #4486. 在每个样品中免疫沉淀的DNA量表示为相当于加入染色质总量，相当于一个。 | |
Western blot analysis of extracts from ER-positive cell lines (MCF7, T-47D, ZR-75-1) and ER-negative cell lines (SK-BR-3 and MCF 10A) using Estrogen Receptor α (D8H8) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).Western blot 方法检测ER-阳性细胞系（MCF7, T-47D, ZR-75-1）提取物和ER-阴性细胞系(SK-BR-3和MCF 10A)，使用的抗体为Estrogen Receptor α (D8H8) Rabbit mAb (上图)或 β-Actin (D6A8) Rabbit mAb #8457 (下图). | |
Confocal immunofluorescent analysis of MCF7 (left) or SK-BR-3 (right) cells using Estrogen Receptor α (D8H8) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).激光共聚焦免疫荧光方法检测MCF7 细胞(左图)或SK-BR-3 细胞(右图)，使用的抗体为 Estrogen Receptor α (D8H8) Rabbit mAb (绿色).肌动蛋白丝用DY-554鬼笔环肽标记（红色）。 |
