货号 | 12534S |
反应种属 | Human/Mouse/Rat/Monkey |
来源宿主 | Rabbit |
应用 | W/IP/ChIP |
目标/特异性 | Smad5 (D4G2) Rabbit mAb recognizes endogenous levels of total Smad5 protein. |
使用方法 | WB(1:1000) IP (1:50) ChIP (1:100) |
供应商 | CST |
背景 | Bone morphogenetic proteins (BMPs) constitute a large family of signaling molecules that regulate a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation, and apoptosis (1,2). BMP receptors are members of the TGF-β family of Ser/Thr kinase receptors. Ligand binding induces multimerization, autophosphorylation, and activation of these receptors (3-5). They subsequently phosphorylate Smad1 at Ser463 and Ser465 in the carboxy-terminal motif SSXS, as well as Smad5 and Smad8 at their corresponding sites. These phosphorylated Smads dimerize with the coactivating Smad4 and translocate to the nucleus, where they stimulate transcription of target genes (5). MAP kinases and CDKs 8 and 9 phosphorylate residues in the linker region of Smad1, including Ser206. The phosphorylation of Ser206 recruits Smurf1 to the linker region and leads to the degradation of Smad1 (6). Phosphorylation of this site also promotes Smad1 transcriptional action by recruiting YAP to the linker region (7). |
存放说明 | -20C |
计算分子量 | 60 |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells treated with Human BMP2 #4697 (50 ng/ml, 1 hr) and either 5 μl of Smad5 (D4G2) 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 ID1 Promoter Primers #5139, human Smad6 promoter 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 (Magnetic Beads) #9003对使用了人BMP2 #4697 (50 ng/ml, 1 hr)和5 μl Smad5 (D4G2) Rabbit mAb兔单抗或2μl 正常Rabbit IgG #2729与4 x 106MCF7交联后进行染色质免疫共沉淀实验。富集的DNA使用SimpleChIP® Human ID1 Promoter 引物 #5139, human Smad6 promoter 引物, 和SimpleChIP® Human α Satellite Repeat 引物 #4486进行实时荧光定量PCR实验。各样品沉淀得到的DNA量比对input染色质总量进行相对定量,每个Input染色质量设定值为1。 | |
Western blot analysis of extracts from various cell lines using Smad5 (D4G2) Rabbit mAb. 使用Smad5 (D4G2)Rabbit mAb兔单抗对多种细胞提取物进行western blot分析。 | |
Immunoprecipitation of Smad5 from HT-1080 cell extracts using Normal Rabbit IgG #2729 (lane 2) or Smad5 (D4G2) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Smad5 (D4G2) Rabbit mAb. 使用正常Rabbit IgG #2729 (lane 2)或Smad5 (D4G2)Rabbit mAb(lane3)对HT-1080细胞提取物进行Smad5蛋白的免疫沉淀实验。Smad5 (D4G2)Rabbit mAb用于western blot分析。 |