Smad3 (phospho S423 + S425) antibody (ab51451)

Product Name 

Smad3 (phospho S423 + S425) antibody

Product type 

Primary antibodies

Description 

Rabbit polyclonal to Smad3 (phospho S423 + S425)

Immunogen 

Synthetic peptide: PSIRCSSpV S_{pBLAST 'PSIRCSSVS' with or}

Reacts with 

Hu, Chk

Predicted to react 
(species key)

Ms, Rat, Cow, Pig, Xl, Zfsh
(Due to sequence homology)

Specificity 

ab51451 is specific for the phosphorylated form of the protein although minimal reactivity with nonphosphorylated human Smad3 can be observed by ELISA and WB. ab51451 also reacts with phosphorylated Smad1 and Smad5 but negligible reactivity is seen against other phosphorylated Smad family members.

Tested applications 
(see key)

ELISA, ICC/IF, WB

Abreviews 

Customer reviews (see key)

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Application notes 
(see key)

Recommended dilutions
ELISA: 1/25000 - 1/100000.
ICC/IF: Use at an assay dependent dilution.
WB: 1/2000 - 1/20000. Predicted molecular weight: 48 kDa. Stimulation with 2 ng/ml TGF-beta for 1 hour is suggested.

Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.

Cellular localization 

In the cytoplasm in the absence of ligand; migration to the nucleus when complexed with SMAD4.

Research areas 

Cancer >> Cancer Metabolism >> Response to hypoxia
Cancer >> Signal transduction >> Nuclear signaling >> SMAD family
Stem Cells >> Signaling Pathways >> TGF beta >> Cytoplasmic
Chromatin and Nuclear Signaling >> Nuclear Signaling Pathways >> SMADs
Signal Transduction >> Signaling Pathway >> Nuclear Signaling >> SMADs

Relevance 

Smad3 is a 50 kDa member of a family of proteins that act as key mediators of TGF beta superfamily signaling in cell proliferation, differentiation and development. The Smad family is divided into three subclasses: receptor regulated Smads, activin/TGF beta receptor regulated (Smad2 and 3) or BMP receptor regulated (Smad 1, 5, and 8); the common partner, (Smad4) that functions via its interaction to the various Smads; and the inhibitory Smads, (Smad6 and 7). Activated Smad3 oligomerizes with Smad4 upon TGF beta stimulation and translocates as a complex into the nucleus, allowing its binding to DNA and transcription factors. Phosphorylation of the two TGF beta dependent serines 423 and 425 in the C terminus of Smad3 is critical for Smad3 transcriptional activity and TGF beta signaling.

Database links 

Raised in 

Rabbit

Clonality 

Polyclonal

Isotype 

IgG

Purity 

Immunogen affinity purified

Storage buffer 

Preservative: 0.01% Sodium Azide
Constituents: 0.15M Sodium chloride, 0.02M Potassium phosphate, pH 7.2

Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS

Purification notes 

ab51451 was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide.

Form 

Liquid

Concentration 

Batch dependent within range: 1.00 - 1.13 mg/ml

Concentration of lot no.     is  

Concentration not available for this lot.

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Storage instructions 

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

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Smad3 (phospho S423 + S425) antibody images:

References for Smad3 (phospho S423 + S425) antibody (ab51451)

  This product has been used in:

Lorda-Diez CI et al. Transforming Growth Factors {beta} Coordinate Cartilage and Tendon Differentiation in the Developing Limb Mesenchyme. J Biol Chem 284:29988-96 (2009). ICC/IF; Chicken. PubMed: 19717568

If you publish research using ab51451 please let us know so that we can cite the reference on this datasheet.

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