| 货号 | 9351L |
| 反应种属 | Human/Mouse |
| 来源宿主 | Rabbit |
| 应用 | W/F/ChIP |
| 目标/特异性 | Phospho-Stat5 (Tyr694) Antibody detects endogenous levelsof Stat5a only when phosphorylated at Tyr694 and Stat5b when phosphorylated at Tyr699. The antibody does not cross-react with the corresponding phospho-tyrosine residues of other Stat proteins, but does cross-react with EGFR. |
| 使用方法 | WB(1:1000) F (1:400) ChIP (1:50) |
| 供应商 | CST |
| 背景 | Stat5 is activated in response to a wide variety of ligands including IL-2, GM-CSF, growth hormone and prolactin. Phosphorylation at Tyr694 is obligatory for Stat5 activation (1,2). This phosphorylation is mediated by Src upon erythropoietin stimulation (3). Stat5 is constitutively active in some leukemic cell types (4). Phosphorylated Stat5 is found in some endothelial cells treated with IL-3, which suggests its involvement in angiogenesis and cell motility (5). Stat5a and Stat5b are independently regulated and activated in various cell types. For instance, interferon treatment predominantly activates Stat5a in U-937 cells and Stat5b in HeLa cells (6). |
| 存放说明 | -20C |
| 计算分子量 | 90 |
Western blot analysis of extracts from TF-1 cells, treated with GM-CSF (25 ng/ml) for the indicated times, using Phospho-Stat5 (Tyr694) Antibody (upper) or Stat5 antibody (lower).Western免疫印迹。用Phospho-Stat5 (Tyr694) Antibody (上图)或者Stat5 antibody (下图)抗体研究经过GM-CSF (25 ng/ml)处理一定时间的TF-1细胞的细胞提取液。 | |
Flow cytometric analysis of K562 cells, untreated (green) or STI571-treated (blue), using Phospho-Stat5 (Tyr694) Antibody.流式细胞仪研究未经处理(绿色)或经过STI571处理(蓝色) 的K562 细胞。所用抗体为Phospho-Stat5 (Tyr694)。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 BaF3 cells starved of IL-3 for 6 hours followed by induction with IL-3 for 45 minutes, and either 20 μl of Phospho-Stat5 (Tyr694) 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® Mouse CIS Intron 1 Primers #5131, mouse SOCS-3 promoter primers, and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. 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。BaF3细胞培养至4 x 106， 并经IL-3饥饿处理6小时后经IL-3诱导45Min,然后用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003进行免疫沉淀实验,本实验中用20 μl Phospho-Stat5 (Tyr694) 抗体 或2 μl Normal Rabbit IgG #2729 。用using SimpleChIP® Mouse CIS Intron 1 Primers #5131对富集的DNA做real-time PCR,所用引物为mouse SOCS-3 promoter primers和SimpleChIP® Mouse RPL30 Intron 2 Primers #7015。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。 |
