Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

PhosphoPlus® Smad2 (Ser465/467) Antibody Duet  #11958

smad   smad2  


No. Size Price
11958S 1 Kit ( 10 western blots ) ¥6,434.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb #3108 100 µl W, H,M,R,Mi, 60 Rabbit IgG
Smad2 (D43B4) XP® Rabbit mAb #5339 100 µl W,IP,IF-IC,F,ChIP, H,M,R,Mk, 60 Rabbit IgG


PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

Cell Signaling Technology (CST)的PhosphoPlus® Duets提供了一种检测蛋白活化状态的方法。每一个Duet都包含了针对感兴趣靶蛋白的激活状态和总蛋白的抗体。这些抗体是基于其优异的性能而被筛选组成试剂盒。

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or TGF-beta treated HeLa and NIH/3T3 cells, using Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb (upper), or Smad2 Antibody #3102 (lower).

使用 Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb (上),或 Smad2 抗体 #3102 (下),对未处理或TGF-beta处理的Hela 和 NIH/3T3细胞提取物进行Western blot 检测。



Confocal immunofluorescent analysis of NIH/3T3 cells, serum-starved (left) or treated with hTGF-β3 #8425 (right), using Smad2 (D43B4) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

血清饥饿处理(左)或 hTGF-β3 #8425 (右)处理NIH/3T3细胞,使用Smad2 (D43B4) XP® Rabbit mAb (绿色)进行共聚焦免疫荧光分析。肌蛋白丝使用 DY-554 phalloidin (红色)标记。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Smad2 (D43B4) XP® Rabbit mAb.

使用 Smad2 (D43B4) XP® Rabbit mAb兔单抗对多种细胞提取物进行Western blot 检测。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #8425 (7 ng/ml) for 1 h and either 10 μl of Smad2 (D43B4) XP® Rabbit mAb #5339 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.

对染色质交联的4 x 106 HaCaT细胞与人 TGF-β3 #8425 (7 ng/ml) 1 h ,10 μl of Smad2 (D43B4) XP® Rabbit mAb #5339 或 2 μl of Normal Rabbit IgG #2729孵育。使用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003进行染色质免疫沉淀。富集的DNA样品使用SimpleChIP® Human CDKN1A Intron 1 引物 #4669, SimpleChIP® Human ID1 Promoter 引物 #5139, 和 SimpleChIP® Human α Satellite Repeat 引物 #4486进行real-time PCR定量。每个样品中的免疫沉淀的DNA量相对于Input中的染色质量进行相对定量,Input中染色质量设定值为1。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using Smad2 (D43B4) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

使用Smad2 (D43B4) XP® Rabbit mAb (蓝色) 和非特异阴性对照(红色)对Hela细胞进行流式细胞分析。


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结合蛋白为目标调控转录反应。[6-8]

  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.

Application References

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Companion Products

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.

PhosphoPlus is a trademark of Cell Signaling Technology, Inc.

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

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