| 货号 | 11958S |
| 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. |
| 供应商 | CST |
| 背景 | Members of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmit TGF-β signals from the cell surface into the nucleus. Three distinct classes of Smads have been defined: the receptor-regulated Smads (R-Smads), which include Smad1, 2, 3, 5, and 8; the common-mediator Smad (co-Smad), Smad4; and the antagonistic or inhibitory Smads (I-Smads), Smad6 and 7 (1-5). Activated type I receptors associate with specific R-Smads and phosphorylate them on a conserved carboxy terminal SSXS motif. The phosphorylated R-Smad dissociates from the receptor and forms a heteromeric complex with the co-Smad (Smad4), allowing translocation of the complex to the nucleus. Once in the nucleus, Smads can target a variety of DNA binding proteins to regulate transcriptional responses (6-8). |
| 存放说明 | -20C |
| 参考文献 | Heldin, C.H. et al. (1997) Nature 390, 465-71. Attisano, L. and Wrana, J.L. (1998) Curr Opin Cell Biol 10, 188-94. Derynck, R. et al. (1998) Cell 95, 737-40. Massagué, J. (1998) Annu Rev Biochem 67, 753-91. Whitman, M. (1998) Genes Dev 12, 2445-62. Wu, G. et al. (2000) Science 287, 92-7. Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7. Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69. |
Western blot analysis of extracts from untreated or TGF-beta treated HeLa and NIH/3T3 cells, using Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb (upper), or Smad2 Antibody #3102 (lower). 使用 Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb (上),或 Smad2 抗体 #3102 (下)，对未处理或TGF-beta处理的Hela 和 NIH/3T3细胞提取物进行Western blot 检测。 | |
Confocal immunofluorescent analysis of NIH/3T3 cells, serum-starved (left) or treated with hTGF-β3 #8425 (right), using Smad2 (D43B4) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). 血清饥饿处理（左）或 hTGF-β3 #8425 (右)处理NIH/3T3细胞，使用Smad2 (D43B4) XP® Rabbit mAb (绿色)进行共聚焦免疫荧光分析。肌蛋白丝使用 DY-554 phalloidin (红色)标记。 | |
Western blot analysis of extracts from various cell lines using Smad2 (D43B4) XP® Rabbit mAb. | |
使用 Smad2 (D43B4) XP® Rabbit mAb兔单抗对多种细胞提取物进行Western blot 检测。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #8425 (7 ng/ml) for 1 h and either 10 μl of Smad2 (D43B4) XP® Rabbit mAb #5339 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® Human CDKN1A Intron 1 Primers #4669, SimpleChIP® Human ID1 Promoter Primers #5139, 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. 对染色质交联的4 x 106 HaCaT细胞与人 TGF-β3 #8425 (7 ng/ml) 1 h ，10 μl of Smad2 (D43B4) XP® Rabbit mAb #5339 或 2 μl of Normal Rabbit IgG #2729孵育。使用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003进行染色质免疫沉淀。富集的DNA样品使用SimpleChIP® Human CDKN1A Intron 1 引物 #4669, SimpleChIP® Human ID1 Promoter 引物 #5139, 和 SimpleChIP® Human α Satellite Repeat 引物 #4486进行real-time PCR定量。每个样品中的免疫沉淀的DNA量相对于Input中的染色质量进行相对定量，Input中染色质量设定值为1。 |
