Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Smad2/3 (D7G7) XP® Rabbit mAb #8685

chip-seq   smad   smad2   smad23   smad3  

No. Size Price
8685S 100 µl ( 10 western blots ) ¥3,750.00 现货查询 购买询价
8685T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
8685 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 52, 60 Rabbit IgG
IP 1:100
F 1:200
IF-IC 1:1600
ChIP 1:100
ChIP-seq 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, ChIP-seq=Chromatin IP-seq,

Specificity / Sensitivity

Smad2/3 (D7G7) XP® Rabbit mAb recognizes endogenous levels of total Smad2/3 protein.

Smad2/3 (D7G7) XP® Rabbit mAb兔单抗识别内源性的总Smad2/3蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding His198 of human Smad2/3 protein.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using Smad2/3 (D7G7) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).

使用Smad2/3 (D7G7) XP® Rabbit mAb兔单抗(蓝色)和Rabbit (DA1E) mAb IgG XP® 同型对照#3900 (红色)对HeLa细胞进行流式细胞分析。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with hTGF-β3 #8425 (7 ng/ml, 1 hr) and either 10 μl of Smad2/3 (D7G7) XP® 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.

用SimpleChIP® Enzymatic Chromatin IP Kit (磁柱) #9003对经过hTGF-β3 #8425 (7 ng/ml, 1 hr)处理的4 x 106 HaCaT细胞和10μl mad2/3 (D7G7) XP®兔单抗或2μl Normal Rabbit IgG #2729进行染色质免疫共沉淀。富集到的DNA与和SimpleChIP® Human CDKN1A Intron1 引物#4669,SimpleChIP® Human ID1 Promoter引物和SimpleChIP®人αSatellite Repeat引物#4486进行荧光实时PCR定量。各样品沉淀得到的DNA量对比input染色质总量进行相对定量,Input中染色质量设定值为1。



Confocal immunofluorescent analysis of HeLa cells, serum starved (left), treated with hTGF-β3 #8425 (100 ng/ml, 30 min, center), or treated with hTGF-β3 and SB43152 (10 ug/mL, 1 hr, right), using Smad2/3 (D7G7) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

用Smad2/3 (D7G7) XP® Rabbit mAb兔单抗 (绿色) 对血清饥饿处理(左),hTGF-β3 #8425 (100 ng/ml, 30 min, 中)处理或hTGF-β3 和SB43152 (10 ug/mL, 1 hr, 右)的HeLa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。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Smad2/3 (D7G7) XP® Rabbit mAb.

用Smad2/3 (D7G7) XP® Rabbit mAb兔单抗对多种细胞提取物进行western blot分析。

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with hTGF-β3 #8425 (7 ng/ml, 1 hr) and 10 μl of Smad2/3 (D7G7) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. The figure shows binding across ID1, a known target gene of Smad2/3 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.


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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


Companion Products

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.

Illumina is a registered trademark of Illumina, Inc.

NEBNext is a registered trademark of New England Biolabs, Inc.

Ultra is a trademark of New England Biolabs, Inc.

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

用户评论 --- 共 0


我要参与评论 :