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Western blot - Smad3 (phospho S423 + S425) antibody (ab51451)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Smad3 (phospho S423 + S425) antibody (ab51451)

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Product Name
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Smad3 (phospho S423 + S425) antibody
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See all Smad3 antibodies (17)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Smad3 (phospho S423 + S425)
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Immunogen
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Synthetic peptide: PSIRCSSpV SpBLAST 'PSIRCSSVS' with or 
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Reacts with
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Hu, Chk
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Predicted to react
(species key)
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Ms, Rat, Cow, Pig, Xl, Zfsh (Due to sequence homology)
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Specificity
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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.
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Tested applications
(see key)
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ELISA, ICC/IF, WB
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Abreviews
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Application notes
(see key)
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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.
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Cellular localization
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In the cytoplasm in the absence of ligand; migration to the nucleus when complexed with SMAD4.
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Research areas
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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
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Relevance
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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.
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Database links
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The links below go to external sites and will open in a new browser window
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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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
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Purification notes
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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.
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Form
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Liquid
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Concentration
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Batch dependent within range: 1.00 - 1.13 mg/ml
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Storage instructions
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Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Smad3 (phospho S423 + S425) antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Smad3 (phospho S423 + S425) antibody images, references, products related to ab51451 and other tools.
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Smad3 (phospho S423 + S425) antibody images:
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Western blot - Smad3 (phospho S423 + S425) antibody (ab51451)
All lanes : Smad3 (phospho S423 + S425) antibody (ab51451) at 1/2000 dilution
Lane 1 : MEF lysate (control) Lane 2 : MEF lysate, after stimulation with 2 ng/ml of TGF-ß for 1 hour Lane 3 : MEF-/-Smad3 lysate (control) Lane 4 : MEF-/-Smad3 lysate, after stimulation with 2 ng/ml of TGF-ß for one hour Lane 5 : Hep3B lysate (control) Lane 6 : Hep3B lysate, after stimulation with 2 ng/ml of TGF-ß for one hour Lane 7 : HEK 293 lysate (control)
Predicted band size : 48 kDa
Image courtesy of Ying Zhang, NIH, CCR, Bethesda, MD.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Smad3 (phospho S423 + S425) antibody (ab51451)
ab51451 at 2.5 µg/ml staining epidermal keratinocytes at 40X. This image shows strong nuclear staining with localization of the antibody as the precipated red siganl and a hematoxylin purple nuclear counterstain.
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References for Smad3 (phospho S423 + S425) antibody (ab51451)
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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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Search PubMed (MEDLINE) for references to Smad3
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Smad3 (phospho S423 + S425) antibody - more information
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Smad3 (phospho S423 + S425) antibody - other tools:
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