Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Smad1 (D59D7) XP® Rabbit mAb #6944

smad   smad1  

No. Size Price
6944S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
6944T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
6944 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Monkey, Endogenous 60 Rabbit IgG
IP 1:100
F 1:100
IF-IC 1:200
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: Xenopus, Bovine,

Specificity / Sensitivity

Smad1 (D59D7) XP® Rabbit mAb recognizes endogenous levels of total Smad1 protein.

Smad1 (D59D7) XP® Rabbit mAb兔单抗识别内源性的总Smad1蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser190 of human Smad1 protein.

合成与人Smad1蛋白邻近190位丝氨酸的氨基酸残基序列一致的肽段,免疫动物获得单克隆抗体。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Smad1 (D59D7) XP® Rabbit mAb.

使用Smad1 (D59D7) XP® Rabbit mAb兔单抗对多种细胞提取物进行western blot实验。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HT-1080 cells using Smad1 (D59D7) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

使用Smad1 (D59D7) XP® Rabbit mAb兔单抗(蓝色)对HT-1080细胞进行流式细胞分析,使用非特异抗体(红色)做为阴性对照。

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) for one hour and either 5 μl of Smad1 (D59D7) XP® 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 (磁珠) #9003将经过人BMP2#4697 (50 ng/ml)处理1小时后的4 x 106 MCF7细胞与5 μl Smad1 (D59D7) XP® Rabbit mAb兔单抗或2μl Normal Rabbit IgG#2729进行免疫共沉淀。富集到的DNA与SimpleChIP®人ID1启动子引物#5139,人SMAD6启动子引物和SimpleChIP® 人αSatellite 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中的染色质量设定值为1。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HT-1080 cells, untreated (left) or treated with human BMP2 #4697 (right), using Smad1 (D59D7) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

未处理(左)或经人BMP2 #4697 处理(右)的HT-1080细胞,使用Smad1 (D59D7) 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。

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).

骨形成蛋白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广泛重要过程[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]。它们随后磷酸化Smad1蛋白的C-末端SSXS模体中的463位丝氨酸和465位丝氨酸,且磷酸化Smad5和Smad8中相应的位置。这些磷酸化的Smad与共激活的Smad4形成二聚体,转移到核内激活靶基因的转录[5]。促分裂原活化蛋白激酶MAPK和周期蛋白依赖性蛋白激酶8和9在Smad1的连接区域磷酸化相应残基包括丝氨酸206。进而募集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.

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

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

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