Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Smad5 (D4G2) Rabbit mAb #12534

smad   smad5  

No. Size Price
12534S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
12534 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 60 Rabbit IgG
IP 1:50
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP,

Specificity / Sensitivity

Smad5 (D4G2) Rabbit mAb recognizes endogenous levels of total Smad5 protein.

Smad5 (D4G2)Rabbit mAb兔单抗可以识别内源性的总Smad5蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro249 of human Smad5 protein.

此单克隆抗体由合成肽段免疫动物产生,该肽段与人Smad5蛋白的Pro249邻近氨基酸残基序列一致。

Chromatin IP

Chromatin IP

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 106 MCF7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 Blotting

Western Blotting

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。

IP

IP

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。

Background

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).

骨形态发生蛋白(BMPs)由一大类信号分子组成,这些分子调控包括形态发生、细胞命运定向、增殖、分化以及凋亡在内的多种关键过程[1,2]。BMP 受体为TGF- β家族丝氨酸/苏氨酸受体成员。配基的结合诱导多聚化、自磷酸化和相应受体的激活[3-5]。它们随后在Smad1蛋白的C-末端SSXS基序磷酸化463位丝氨酸和465位丝氨酸,并且也磷酸化Smad5和Smad8相应的位置。这些磷酸化的Smad与共激活的Smad4形成二聚体,转移到核内刺激靶基因的转录[5]。促分裂原活化蛋白激酶 MAPK和周期蛋白依赖性蛋白激酶CDKs 8和9在磷酸化Smad1的包括丝氨酸206等氨基酸残基在内的连接区域。丝氨酸206的磷酸化召集Smurf1到接头区域导致Smad1的降解[6]。这一位点的磷酸化也招募YAP到接头区域促进Smad1的转录激活[7]。

  1. Hogan, B.L. (1996) Genes Dev 10, 1580-94.
  2. Hoodless, P.A. et al. (1996) Cell 85, 489-500.
  3. Klemm, J.D. et al. (1998) Annu Rev Immunol 16, 569-92.
  4. Kretzschmar, M. et al. (1997) Genes Dev 11, 984-95.
  5. Whitman, M. (1998) Genes Dev 12, 2445-62.
  6. Sapkota, G. et al. (2007) Mol Cell 25, 441-54.
  7. Alarcón, C. et al. (2009) Cell 139, 757-69.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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