Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad9 (Ser465/467) (D5B10) Rabbit mAb #13820

sc-12353   Smad   Smad1   Smad158   Smad159   Smad5   Smad8   Smad9  

No. Size Price
13820S 100 µl ( 10 western blots ) ¥4,050.00 现货查询 购买询价 防伪查询
13820T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价 防伪查询
13820 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 60 Rabbit IgG
IP 1:50
F 1:400
IF-IC 1:800

Species cross-reactivity is determined by western blot.

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


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

Specificity / Sensitivity

Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb recognizes endogenous levels of Smad1 and Smad5 protein when phosphorylated at Ser463/465. This antibody is also predicted to detect Smad9 (Smad8) when phosphorylated at Ser465/467.

Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb 兔单抗可以识别内源性的磷酸化Smad1和Smad5 蛋白,但是需要er463和Ser465,被双重磷酸化,抗体也可能识别Ser465/467磷酸化的Smad9 (Smad8)蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser463/465 of human Smad1 and Smad5 protein.


Western Blotting

Western Blotting

Western blot analysis of extracts from Hep G2 or MEF cells, untreated (-) or treated with Human BMP2 #4697 (50 ng/ml, 30 min; +), using Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb (upper) and Smad1 (D59D7) XP® Rabbit mAb #6944 (lower).

使用Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb兔单抗 (上) and Smad1 (D59D7) XP® Rabbit mAb 兔单抗#6944 (下),对Human BMP2 #4697 (50 ng/ml, 30 min; +)处理或无处理(-)的Hep G2 或 MEF细胞提取物进行Western blot分析。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HT-1080 cells, untreated (blue) or treated with Human BMP2 #4697 (green), using Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

使用Phospho-Smad1/5 (Ser463/465) (D5B10) Rabbit mAb兔单抗对Human BMP2 #4697 (绿色)处理或无处理(蓝色)的HT-1080细胞进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 偶联) #4412做为二抗。



Confocal immunofluorescent analysis of HT-1080 cells, serum-starved (overnight; left) or serum-starved and treated with Human BMP2 #4697 (50 ng/ml, 30 min; right), using Phospho-Smad1/5 (Ser463/Ser465) (D5B10) Rabbit mAb (green) and β-Actin (13E5) Rabbit mAb (Alexa Fluor® 647 Conjugate) #8584 (red). Blue pseudocolor = Propidium Iodide (PI)/RNase Staining Solution #4087.

血清饥饿(过夜处理,左)或血清饥饿和Human BMP2 #4697 (50 ng/ml, 30 min; 右)处理的HT-1080细胞,使用Phospho-Smad1/5 (Ser463/Ser465) (D5B10) Rabbit mAb兔单抗 (绿色) 和 β-Actin (13E5) Rabbit mAb 兔单抗(Alexa Fluor® 647 偶联) #8584 (红色)进行共聚焦免疫荧光分析,蓝色伪彩=Propidium Iodide (PI)/RNase 染色液 #4087。


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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