Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Smad3 (C67H9) Rabbit mAb #9523

sc-101154   smad   smad3  

No. Size Price
9523S 100 µl ( 10 western blots ) ¥3,250.00 现货查询 购买询价 防伪查询
9523T 20 µl ( 2 western blots ) ¥1,200.00 现货查询 购买询价 防伪查询
9523 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 52 Rabbit IgG
IP 1:100
F 1:100
IF-IC 1:100
ChIP 1:50

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,


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

Specificity / Sensitivity

Smad3 (C67H9) Rabbit mAb detects endogenous levels of total Smad3 protein. No cross reactivity was detected with other family members.

Smad3 (C67H9) Rabbit mAb兔单抗可以识别内源的总Smad3蛋白。经检测与其他家族成员没有交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues at the amino terminus of Smad3.


Western Blotting

Western Blotting

Western blot analysis of extracts from HT1080 (human), C2C12 (mouse) and B35 (rat) using Smad3 (C67H9) Rabbit mAb.

使用Smad3 (C67H9) Rabbit mAb兔单抗对HT1080 (人), C2C12 (小鼠)和B35 (大鼠)细胞提取物进行western blot分析。

Western Blotting

Western Blotting

Western blot analysis of extracts from HT1080 cells, treated with TGF-β1, TGFR inhibitor SB-431542 or BMP-2, using Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb #9520 (upper) or total Smad3 (C67H9) Rabbit mAb #9523 (lower).

使用Phospho-Smad3 (Ser423/425) (C25A9)Rabbit mAb兔单抗#9520 (上)或 total Smad3 (C67H9) Rabbit mAb兔单抗 #9523 (下)对经过 TGF-β1, TGFR 抑制因子 SB-431542 或BMP-2处理后的HT1080细胞提取物进行western blot分析。



Confocal immunofluorescent analysis of HT1080 cells, untreated (left) or TGFβ-treated (right), using Smad3 (C67H9) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

使用Smad3 (C67H9) 兔单抗(绿色)对处理(左)或 TGFβ-处理(右)的HT1080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。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HT-1080 cells using Smad3 (C67H9) Rabbit mAb #9523 (blue) compared to a nonspecific negative control antibody (red).

使用Smad3 (C67H9) Rabbit mAb兔单抗 #9523 对Ht-1080细胞进行流式细胞分析,使用非特异性抗体做为阴性对照(红色)。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #3706 (7 ng/ml) for 1 h and either 10 μl of Smad3 (C67H9) 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 CDKN1A Intron 1 Primers #4669, SimpleChIP® Human ID1 Promoter Primers #5139, 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对经过人TGF-β3 #3706 (7 ng/ml)处理的4 x 106 HaCaT细胞和10μl Smad3 (C67H9)Rabbit mAb兔单抗或2 μlNormal Rabbit IgG#2729进行染色质免疫共沉淀。富集到的DNA使用人CDKN1A内含子1引物#4669, SimpleChIP® 人ID1启动子引物#5139,人SMAD6启动子引物和SimpleChIP®人α Satellite Repeat引物#4486进行荧光实时PCR定量。各样品沉淀得到的DNA量比对于nput染色质量进行相对定量,Input的染色质量设定值为1。


Members of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmit TGF-β signals from the cell surface into the nucleus. Three distinct classes of Smads have been defined: the receptor-regulated Smads (R-Smads), which include Smad1, 2, 3, 5, and 8; the common-mediator Smad (co-Smad), Smad4; and the antagonistic or inhibitory Smads (I-Smads), Smad6 and 7 (1-5). Activated type I receptors associate with specific R-Smads and phosphorylate them on a conserved carboxy terminal SSXS motif. The phosphorylated R-Smad dissociates from the receptor and forms a heteromeric complex with the co-Smad (Smad4), allowing translocation of the complex to the nucleus. Once in the nucleus, Smads can target a variety of DNA binding proteins to regulate transcriptional responses (6-8).

Smad信号转导分子家族成员是胞内通路中TGF-β信号从细胞表面传送到细胞核的重要组成部分。Smad 可被分为三类:受体调控型Smad(简称R-Smad),主要包括Smad1,2,3,5和8;共同介导型Smad(co-Smad)包括Smad4;还有拮抗型或称抑制型Smad(I-Smads),包括Smad6和Smad7[1-5]。激活的I型受体与特异的R-Smad有关,并能在R-Smad的保守的C端SSXS基序处对其磷酸化。磷酸化的R-Smad与受体分离,再与co-Smad(Smad4)形成异侧复合物,使复合物迁移入核。一旦进入核内,Smad可以各种DNA绑定蛋白为目标调控转录反应。[6-8]

Following stimulation by TGF-β, Smad2 and Smad3 become phosphorylated at their carboxyl termini (Ser465 and 467 on Smad2; Ser423 and 425 on Smad3) by TGF-β Receptor I. Phosphorylated Smad 2/3 can complex with Smad4, translocate to the nucleus and regulate gene expression (9-11).

TGF-β刺激后,Smad2和Smad3羧基末端丝氨酸残基(Smad2的465位和467位丝氨酸,Smad3的423和425位丝氨酸)被TGF-β Receptor 1磷酸化。磷酸化的Smad2/3与Smad4形成复合物进入细胞核调控基因表达(9-11)。

  1. Heldin, C.H. et al. (1997) Nature 390, 465-71.
  2. Attisano, L. and Wrana, J.L. (1998) Curr Opin Cell Biol 10, 188-94.
  3. Derynck, R. et al. (1998) Cell 95, 737-40.
  4. Massagué, J. (1998) Annu Rev Biochem 67, 753-91.
  5. Whitman, M. (1998) Genes Dev 12, 2445-62.
  6. Wu, G. et al. (2000) Science 287, 92-7.
  7. Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7.
  8. Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69.
  9. Abdollah, S. et al. (1997) J. Biol. Chem. 272, 27678-27685.
  10. Souchelnytskyi, S. et al. (1997) J. Biol. Chem. 272, 28107-28115.
  11. Liu, X. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 10669-10674.

Application References

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

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

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