| 货号 | 5339S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/F/ChIP |
| 使用方法 | WB(1:1000) IP (1:50) F (1:200) IF-IC (1:100) ChIP (1:100) |
| 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 |
| 计算分子量 | 60 |
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兔单抗（绿色）进行激光共聚焦荧光分析。使用DY-554 phalloidin（红色）标记肌动蛋白丝。 | |
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分析。 | |
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. 用人TGF-β3 #8425 (7 ng/ml)对4 x 106 HaCaT细胞处理 1小时后获取交联染色质，使用SimpleChIP® Enzymatic Chromatin IP Kit (磁柱) #9003 的10 μl Smad2 (D43B4) XP® Rabbit mAb兔单抗#5339或2μl Normal Rabbit IgG #2729进行染色质免疫共沉淀。富集到的DNA随后使用SimpleChIP®人CDKN1A内含子1引物#4669，SimpleChIP®人ID1启动子引物#5139和SimpleChIP® Human α Satellite Repeat 引物#4486进行荧光实时PCR定量。各样品沉淀得到的DNA量通过对比input染色质总量进行相对定量，Input的染色质量设定值为1。 | |
Flow cytometric analysis of HeLa cells using Smad2 (D43B4) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red). 使用Smad2 (D43B4) XP® Rabbit mAb兔单抗（蓝色）对HeLa细胞进行流式细胞分析，采用非特异抗体做为阴性对照（红色）。 |
