| 货号 | 7649S |
| 反应种属 | Human/Mouse/Rat |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/F/ChIP |
| 目标/特异性 | Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb recognizes endogenous levels of Stat1 protein only when phosphorylated at Tyr701. |
| 使用方法 | WB(1:1000) IP (1:50) F (1:200) IF-IC (1:50) ChIP (1:100) |
| 供应商 | CST |
| 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 |
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with hIFN-α1 #8927 (100 ng/mL, 30 min; right), using Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red).共聚焦免疫荧光分析HeLa细胞，未经处理 (左图) 和经 hIFN-α1 #8927 (100 ng/mL, 30 min; 右图)处理，所用抗体为Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb (绿色)和β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (红色)。 | |
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with hIFN-α1 #8927 (green), using Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb.流式细胞仪分析Jurkat细胞未经处理 (蓝色)或经过hIFN-α1 #8927 处理(绿色)，所用抗体为Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb。 | |
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 5 μl of Phospho-Stat1 (Tyr701) (D4A7) 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 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) 处理30分钟,然后用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003进行免疫沉淀实验,本实验中用5 μl Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb 或2 μl Normal Rabbit IgG #2729 。用人的IRF-1启动子引物和SimpleChIP® Human TAP1 Promoter Primers #5148，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, A20, and PC-12 cells, untreated or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 30 min), using Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb (upper) or Stat1 Antibody #9172 (lower).Western免疫印迹。用Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb (上图) 或者 Stat1 Antibody #9172 (下图)研究未经处理的和经hIFN-α1 #8927 (10 ng/ml, 30 min)处理的 HeLa, A20和PC-12 细胞的细胞提取液。 |
