| 货号 | 8215S |
| 描述 | PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CSTs product offering based upon superior performance in specified applications.CST公司生产的PhosphoPlus® Duets为蛋白的活性形式研究提供了方法。每一个Duet 包含了目标蛋白的活性形式和总蛋白。这些抗体是从CST生产的在特定的实验中有高性能的产品中挑选的。 |
| 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).Stat5的激活是通过一系列的配体的结合而完成，包括 IL-2, GM-CSF,生长激素和泌乳刺激素。磷酸化Tyr694位点对 Stat5的活性是必须的(1,2)。此位点的磷酸化是在红细胞生成素的作用下由Src介导而完成(3)。 Stat5在某些白血病类型的细胞中有组成性的活性(4)。用IL-3处理的内皮细胞中有磷酸化的Stat5，这就表明它参与了血管再生和细胞的运动(5)。在不同细胞类型中 Stat5a 和 Stat5b 是被独立调控和激活的。例如,干扰素处理后在U-937细胞中Stat5a的活性较强但在HeLa细胞中Stat5b的活性较强(6)。 |
| 存放说明 | -20C |
| 参考文献 | Gouilleux, F. et al. (1994) EMBO J 13, 4361-9. Wakao, H. et al. (1994) EMBO J 13, 2182-91. Okutani, Y. et al. (2001) Oncogene 20, 6643-50. Demoulin, J.B. et al. (1999) J Biol Chem 274, 25855-61. Dentelli, P. et al. (1999) J Immunol 163, 2151-9. Meinke, A. et al. (1996) Mol Cell Biol 16, 6937-44. |
Western blot analysis of extracts from various cell lines using the Stat5 Antibody. | |
Western blot analysis of extracts from HeLa, L929 and PC12 cells, using Stat5 (3H7) Rabbit mAb. | |
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 10 μl of Stat5 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 SOCS3 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. | |
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 Stat5 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. | |
Western blot analysis of extracts from UT-7 cells, untreated or treated with erythropoietin (EPO; 3 units/ml for 5 min), TF-1 cells, untreated or treated with Human Granulocyte Macrophage Colony Stimulating Factor #8922 (hGM-CSF; 100ng/ml for 10 min), and NK-92 cells, untreated or treated with Human Interleukin-2 #8907 (hIL-2; 100ng/ml for 10 min), using Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb (upper) or total Stat5 (3H7) Rabbit mAb #9358 (lower). | |
Flow cytometric analysis of TF-1 cells, untreated (blue) or GM-CSF treated (green), using Phospho-Stat5 (Tyr694) (D47E7) XP® Rabbit mAb. | |
Confocal immunofluorescent analysis of A-431 cells, EGF-treated (left) or untreated (right), using Phospho-Stat5 (Tyr694) XP®(D47E7) Rabbit mAb (green) and Pan-Keratin (C11) Mouse mAb #4545 (red). |
