| 货号 | 9520S |
| 反应种属 | Human/Mouse/Rat |
| 来源宿主 | Rabbit |
| 应用 | W/IP/ChIP |
| 目标/特异性 | Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb detects endogenous levels of Smad3 when phosphorylated at Ser423/425. This antibody does not cross-react reactivity with other family members. |
| 使用方法 | WB(1:1000) IP (1:50) 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 | 52 |
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 5 μl of Phospho-Smad3 (Ser423/425) (C25A9) 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, human c-Myc intron 1 primers, 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 (磁珠偶联) #9003对经过人TGF-β3 #3706 (7 ng/ml)处理1小时的4 x 106 HaCaT细胞和5μl Phospho-Smad3 (Ser423/425) (C25A9)兔单抗或2μlNormal 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c-Myc内含子引物和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input染色质总量进行相对定量，Input染色质量设定值为1。 | |
Western blot analysis of extracts from HT-1080, C2C12, or KNRK cells, untreated (-) or treated with TGF-β (10 ng/ml, 30 min; +), using Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb (upper) or total Smad3 (C67H9) Rabbit mAb #9523 (lower). |
