| 货号 | 9172T |
| 反应种属 | Human,Mouse,Rat,Monkey, |
| 来源宿主 | Rabbit |
| 应用 | WB, IP , ChIP |
| 目标/特异性 | Stat1 Antibody detects endogenous levels of total Stat1 protein. The antibody detects both Stat1alpha (91kDa) and Stat1beta (84 kDa) isoforms. |
| 使用方法 | WB(1:1000) IP (1:100) ChIP (1:100) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | The Stat1 transcription factor is activated in response to a large number of ligands (1) and is essential for responsiveness to IFN-α and IFN-γ (2,3). Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization, nuclear translocation, and DNA binding (4). Stat1 protein exists as a pair of isoforms, Stat1α (91 kDa) and the splice variant Stat1β (84 kDa). In most cells, both isoforms are activated by IFN-α, but only Stat1α is activated by IFN-γ. The inappropriate activation of Stat1 occurs in many tumors (5). In addition to tyrosine phosphorylation, Stat1 is also phosphorylated at Ser727 through a p38 mitogen-activated protein kinase (MAPK)-dependent pathway in response to IFN-α and other cellular stresses (6). Serine phosphorylation may be required for the maximal induction of Stat1-mediated gene activation. |
| 存放说明 | -20C |
| 计算分子量 | 84, 91 |
| 参考文献 | 1 . Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120. 2 . Durbin, J.E. et al. (1996) Cell 84, 443-50. 3 . Meraz, M.A. et al. (1996) Cell 84, 431-42. 4 . Ihle, J.N. et al. (1994) Trends Biochem Sci 19, 222-7. 5 . Frank, D.A. (1999) Mol. Med. 5, 432-456. 6 . Wen, Z. et al. (1995) Cell 82, 241-50. |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and either 10 μl of Stat1 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 human IRF-1 promoter primers, SimpleChIP® Human TAP1 Promoter Primers #5148, 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.染色质免疫共沉淀。HT-1080 细胞培养至4 x 106 , 并经IFN-γ (50 ng/ml,30 min) 处理,然后用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003进行免疫沉淀实验,本实验中用10 μl of Stat1 抗体或2 μl Normal Rabbit IgG #2729 抗体。用human IRF-1 promoter primers, SimpleChIP® Human TAP1 Promoter Primers #5148 和SimpleChIP® Human α Satellite Repeat Primers #4486. 对富集的DNA做real-time PCR。每个样本中沉淀的DNA量定义为相对信号与输入的总染色质相比的数值。 | |
Western blot analysis of extracts from HeLa cells 48 hours following mock transfection, transfection with non-targeted (control) siRNA or transfection with Stat1 siRNA. Stat1 was detected using Stat1 Antibody #9172, and p42 was detected using p42 MAPK Antibody #9108. The Stat1 Antibody confirms silencing of Stat1 expression, and the p42 MAPK Antibody is used to control for protein loading and siRNA specificity.Western免疫印迹。HeLa细胞经过对照转染，转染了non-targeted (对照) siRNA ,转染了Stat1 siRNA的质粒48小时后抽提细胞液,用Stat1 Antibody #9172研究Stat1蛋白, 用p42 MAPK Antibody #9108研究 p42 蛋白。Stat1 抗体是为了验证基因沉默靶基因Stat1蛋白的表达, p42 MAPK 抗体是为了蛋白上样的参照和siRNA 的特异性。 | |
Western blot analysis of extracts from SK-MEL-28 cells, untreated or IFN-alpha-treated (100 ng/ml), using Phospho-Stat1 (Tyr701) Antibody #9171 (upper) or Stat1 Antibody (lower).Western免疫印迹。用Phospho-Stat1 (Tyr701) Antibody #9171 (上图) or Stat1 Antibody (下图)研究未经处理的或经 IFN-alpha(100 ng/ml)处理的SK-MEL-28 细胞的细胞提取液。 |
