Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Smad4 (D3M6U) Rabbit mAb #38454

co-Smad  

No. Size Price
38454S 100 µl ( 10 western blots ) ¥3,250.00 现货查询 购买询价 防伪查询
38454T 20 µl ( 2 western blots ) ¥1,200.00 现货查询 购买询价 防伪查询
38454 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 70 Rabbit IgG
IP 1:200
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: Dog, Guinea Pig, Horse,

Specificity / Sensitivity

Smad4 (D3M6U) Rabbit mAb recognizes endogenous levels of total Smad4 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp165 of human SMAD4 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Smad4 (D3M6U) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). HT-29 and COLO 205 are Smad4-null mutant cell lines, confirming specificity of the antibody.

IP

IP

Immunoprecipitation of Smad4 protein from HCT 116 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Smad4 (D3M6U) Rabbit mAb. Western blot analysis was performed using Smad4 (D3M6U) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with TGF-β1 #8915 (7 ng/mL, 1 hr) and either 5 µl of Smad4 (D3M6U) 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 ID1 Promoter Primers #5139, human JunB promoter primers, 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 (equivalent to one).

Background

Members of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmits TGF-β signals from the cell surface into the nucleus. Three distinct classes of Smads have been defined: the recepter-regulated Smads (R-Smads), which include Smad1, 2, 3, 5, 8; the common-mediator Smad (co-Smad), Smad4; and the antagonistic or inhibitory Smads (I-Smads), Smad6 and 7 (1-5). Briefly, activated type I receptors associate with specific R-Smads and phosphorylate them on a conserved SSXS motif at the carboxy-terminus of the proteins. The phosphorylated R-Smad dissociates from the receptor and forms a heteromeric complex with the co-Smad, Smad4, and together the complex moves to the nucleus. Once in the nucleus, Smads can target a variety of DNA binding proteins to regulate transcriptional responses (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. Massague, J. (1998) Annu. Rev. Biochem. 67, 753-91.
  5. Whitman, . (1998) Genes Dev. 12, 2445-62.
  6. Wrana, J. (2000) Science 23, 1-9.
  7. Attisano, L. and Wrana, J. (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!

Protocols

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.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

Tween is a registered trademark of ICI Americas, Inc.

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

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