Product Pathways - TGF-beta/Smad Signaling

Phospho-Smad Antibody Sampler Kit #9963

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Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Source
Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb # 9516	40 microliters	W F	H M	60	Rabbit
Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb # 3108	40 microliters	W	H M Mi	60	Rabbit
Smad2 (86F7) Rabbit mAb # 3122	40 microliters	W IP IF-IC	H Mk	60	Rabbit
Smad5 Antibody # 9517	40 microliters	W	H M R Mk	60	Rabbit
Smad4 Antibody # 9515	40 microliters	W	H M R Mk	70	Rabbit
Smad6 Antibody # 9519	40 microliters	W	H M Mk	62	Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074	100 microliters				Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Mi=Mink

Specificity / Sensitivity

Activation state antibodies detect their intended targets only when phosphorylated at the indicated site. Total Smad 2,4,5 and 6 Antibodies detect endogenous levels of total Smad 2, 4, 5 and 6, respectively.

Source / Purification

Rabbit monoclonal antibodies are produced by immunizing rabbits with a synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Ser463/465 of human Smad5, Ser465/467 of human Smad2, and a synthetic peptide (KLH-coupled) corresponding to residues surrounding Trp85 of human Smad2. Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to the residues surrounding Gln252 of human Smad5, Pro278 of human Smad4, and Cys55 of human Smad6. Antibodies are purified by protein A and peptide affinity chromatography.

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 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).

1. Heldin, C.H. et al. (1997) Nature 390, 465-471.
2. Attisano, L. and Wrana, J.L. (1998) Curr. Opin. Cell Biol. 10, 188-194.
3. Derynck, R. et al. (1998) Cell 95, 737-740.
4. Massague, J. (1998) Annu. Rev. Biochem. 67, 753-791.
5. Whitman, M. et al. (1998) Genes Dev. 12, 2445-2462.
6. Wrana, J. (2000) Science 23, 1-9.
7. Attisano, L. and Wrana, J. (2002) Science 296, 1646-1647.
8. Moustakas, A. et al. (2001) J. Cell Sci. 114, 4359-4369.

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

• 9511 Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad8 (Ser426/428) Antibody
• 3101 Phospho-Smad2 (Ser465/467) Antibody
• 3102 Smad2/3 Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.