| 货号 | 4134S |
| 反应种属 | Human |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/F/ChIP |
| 目标/特异性 | Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb recognizes endogenous levels of Stat4 protein only when phosphorylated at Tyr693. |
| 使用方法 | WB(1:1000) IP (1:100) F (1:800) IF-IC (1:800) ChIP (1:50) |
| 供应商 | CST |
| 背景 | The Jak-Stat signaling pathway is utilized by a large number of cytokines, growth factors, and hormones (1). Receptor-mediated tyrosine phosphorylation of Jak family members triggers phosphorylation of Stat proteins, resulting in their nuclear translocation, binding to specific DNA elements, and subsequent activation of transcription. The remarkable range and specificity of responses regulated by the Stats is determined, in part, by the tissue-specific expression of different cytokine receptors, Jaks, and Stats, as well as by the combinatorial coupling of various Stat members to different receptors (2). Stat4 is predominantly expressed in the spleen, thymus, and testis and has been most extensively investigated as the mediator of IL-12 responses (3-8). Activation of Stat4 is associated with phosphorylation at Tyr693 (9). |
| 存放说明 | -20C |
| 计算分子量 | 81 |
Confocal immunofluorescent analysis of NK-92 cells, starved of IL-2 (5 hr) and then either untreated (upper) or IL-12-treated (50 ng/mL, 15 min; lower), using Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).共聚焦免疫荧光分析经IL-2饥饿 5 小时 NK-92细胞，未经处理 (上图) 和进过IL-12(50 ng/mL, 15 min) 处理(下图)，Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb(绿色)和 β-Actin (8H10D10) Mouse mAb #3700(红色)。 | |
Western blot analysis of extracts from NK-92 cells, untreated (-) or treated (+) with Human Interleukin-2 (hIL-2) #8907 (10 ng/ml, 15 min) or human interleukin-12 (hIL-12, 50 ng/ml, 15 min), using Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb (upper) and Stat4 (C46B10) Rabbit mAb #2653 (lower).Western免疫印迹。用Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb (上图) 和Stat4 (C46B10) Rabbit mAb #2653 (下图) 研究未经处理的和经(-) 或 经过 hIL-2 #8907 (10 ng/ml, 15 min) 或者(hIL-12, 50 ng/ml, 15 min)处理(+) 的NK-92 细胞的细胞提取液。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NK-92 cells starved of IL-2 overnight then treated with IL-12 (10 ng/ml, 4 hr) and either 20 μl of Phospho-Stat4 (Tyr693) (D2E4) 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 PRF1 Promoter Primers #9014, 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。NK-92细胞培养至4 x 106，IL-2饥饿过夜并经IL-12 (10 ng/ml)处理4小时,然后用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003进行免疫沉淀实验,所用的抗体为20 μl Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb或2 μl of Normal Rabbit IgG #2729，本实验中用IRF-1 promoter primers, SimpleChIP® Human PRF1 Promoter Primers #9014和 SimpleChIP® Human α Satellite Repeat Primers #4486引物对富集的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。 | |
Flow cytometric analysis of NK-92 cells, untreated (blue) or treated with IL-12 (green), using Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb.流式细胞仪分析NK-92 细胞，未经处理(蓝色) 或经过IL-12处理(绿色) ，所用抗体为Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb。 |
