| 货号 | 9523S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/F/ChIP |
| 目标/特异性 | Smad3 (C67H9) Rabbit mAb detects endogenous levels of total Smad3 protein. No cross reactivity was detected with other family members. |
| 使用方法 | WB(1:1000) IP (1:100) F (1:100) IF-IC (1:100) ChIP (1:50) |
| 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 | 52 |
Western blot analysis of extracts from HT1080 (human), C2C12 (mouse) and B35 (rat) using Smad3 (C67H9) Rabbit mAb. 使用Smad3 (C67H9) Rabbit mAb兔单抗对HT1080 (人), C2C12 (小鼠)和B35 (大鼠)细胞提取物进行western blot分析。 | |
Western blot analysis of extracts from HT1080 cells, treated with TGF-β1, TGFR inhibitor SB-431542 or BMP-2, using Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb #9520 (upper) or total Smad3 (C67H9) Rabbit mAb #9523 (lower). 使用Phospho-Smad3 (Ser423/425) (C25A9)Rabbit mAb兔单抗#9520 (上)或 total Smad3 (C67H9) Rabbit mAb兔单抗 #9523 (下)对经过 TGF-β1, TGFR 抑制因子 SB-431542 或BMP-2处理后的HT1080细胞提取物进行western blot分析。 | |
Confocal immunofluorescent analysis of HT1080 cells, untreated (left) or TGFβ-treated (right), using Smad3 (C67H9) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). 使用Smad3 (C67H9) 兔单抗（绿色）对处理（左）或 TGFβ-处理（右）的HT1080细胞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。 | |
Flow cytometric analysis of HT-1080 cells using Smad3 (C67H9) Rabbit mAb #9523 (blue) compared to a nonspecific negative control antibody (red). 使用Smad3 (C67H9) Rabbit mAb兔单抗 #9523 对Ht-1080细胞进行流式细胞分析，使用非特异性抗体做为阴性对照（红色）。 | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #3706 (7 ng/ml) for 1 h and either 10 μl of Smad3 (C67H9) 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 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. 使用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003对经过人TGF-β3 #3706 (7 ng/ml)处理的4 x 106 HaCaT细胞和10μl Smad3 (C67H9)Rabbit mAb兔单抗或2 μlNormal Rabbit IgG#2729进行染色质免疫共沉淀。富集到的DNA使用人CDKN1A内含子1引物#4669, SimpleChIP®人ID1启动子引物#5139，人SMAD6启动子引物和SimpleChIP®人α Satellite Repeat引物#4486进行荧光实时PCR定量。各样品沉淀得到的DNA量比对于nput染色质量进行相对定量，Input的染色质量设定值为1。 |
