Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Smad2 (D43B4) XP® Rabbit mAb #5339

smad   smad2  

No. Size Price
5339S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
5339T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
5339 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 60 Rabbit IgG
IP 1:50
F 1:200
IF-IC 1:100
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,

Specificity / Sensitivity

Smad2 (D43B4) XP® Rabbit mAb detects endogenous levels of total Smad2 protein. This antibody does not cross-react with Smad3.

Smad2 (D43B4) XP® Rabbit mAb兔单抗可以识别内源性的总Smad2蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of mouse Smad2 protein.

合成与小鼠Smad2蛋白邻近氨基末端的氨基酸残基序列一致的肽段,免疫动物获得单克隆抗体。

IF-IC

IF-IC

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。

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分析。

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.

用人TGF-β3 #8425 (7 ng/ml)对4 x 106 HaCaT细胞处理 1小时后获取交联染色质,使用SimpleChIP® Enzymatic Chromatin IP Kit (磁柱) #9003 的10 μl Smad2 (D43B4) XP® Rabbit mAb兔单抗#5339或2μl Normal Rabbit IgG #2729进行染色质免疫共沉淀。富集到的DNA随后使用SimpleChIP® 人CDKN1A内含子1引物#4669,SimpleChIP® 人ID1启动子引物#5139和SimpleChIP® Human α Satellite Repeat 引物#4486进行荧光实时PCR定量。各样品沉淀得到的DNA量通过对比input染色质总量进行相对定量,Input的染色质量设定值为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兔单抗(蓝色)对HeLa细胞进行流式细胞分析,采用非特异抗体做为阴性对照(红色)。

Background

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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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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