Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb #9516

BMP   MAD   sc-12353   smad   smad1   smad15   smad5  

No. Size Price
9516S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
9516T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
9516 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 60 Rabbit IgG
F 1:400
IF-IC 1:50

Species cross-reactivity is determined by western blot.

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

Specificity / Sensitivity

Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb detects endogenous levels of Smad1/5 only when dually phosphorylated at Ser463 and Ser465. The antibody does not cross-react with other Smad-related proteins.

Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb兔单抗仅能识别463/465位丝氨酸双重磷酸化的内源性Smad1/5蛋白。本抗体与其他Smad相关蛋白没有交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser463/465 of human Smad5.

合成与人源Smad5 蛋白的邻近463/465位丝氨酸氨基酸残基序列一致的磷酸化肽段,免疫动物获得单克隆抗体。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) or BMP-treated (green), using Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb compared to a nonspecific negative control antibody (red).

使用pho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb兔单抗对未处理(蓝色)或BMP处理(绿色)的HeLa细胞进行流式细胞分析,用非特异性抗体做阴性对照(红色)。

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or BMP-4-treated HeLa or NIH/3T3 cells using Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb.

使用 Phospho-Smad1/5 (Ser463/465) (41D10)Rabbit mAb兔单抗对未处理或经过BMP-4处理的HeLa或NIH/3T3细胞提取物进行western blot分析。



Confocal immunofluorescent analysis of HT-1080 cells, BMP-treated (left) and untreated (right), using Phospho-Smad5 (Ser463/Ser465) (41D10) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

使用Phospho-Smad5 (Ser463/Ser465) (41D10) Rabbit mAb兔单抗(绿色)对BMP-处理(左)和未处理(右)的HT-1080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。



Confocal immunofluorescent analysis of HT1080 cells, serum-starved (left) or serum-starved then treated with hBMP2 (50 ng/ml, 30 min; right) using Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb (green) and Cox IV (4D11-B3-E8) Mouse mAb #11967 (red).


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.

U.S. Patent No. 5,675,063.

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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