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A synthetic phospho specific peptide corresponding to residues surrounding Ser423 and Ser425 of human Smad3.
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our Abpromise guarantee covers the use of ab52903 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/2000. Predicted molecular weight: 48 kDa.
Avoid using milk, casein, and phopshorylated proteins in general in the blocking buffer and in the antibody diluent. We recommend a solution of 5% BSA (bovine serum albumin).
|ICC/IF||1/100 - 1/250.|
|ELISA||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration. PubMed: 23667656|
|IHC-P||1/100 - 1/250.|
ab52903 staining Smad3 in mouse primary embryonic epicardial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% formaldehyde, permeabilized with 0.5% Triton X-100 and blocked with PBS + 1% BSA + 10% goat serum + 0.1% Triton X-100 for 1 hour at 20°C. Samples were incubated with primary antibody (1/100 in PBS + 1% BSA + 10% goat serum + 0.1% Triton X-100) for 16 hours at 4°C. An Alexa Fluor®488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This image is a courtesy of Aaron Gardner
ab52903 staining Smad3 (phospho S423 + S425) in human TII Pneumocyte A549c ells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde and permeabilized with 0.1% Triton x100 before blocking with 3% BSA for 1 hour at RT. Samples were incubated with primary antibody (1/200: in 3% BSA in 1x PBST) for 24 hours at 4°C. A TRITC-conjugated goat polyclonal to rabbit IgG was used as secondary antibody at 1/200 dilution.