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Fibronectin was purified from Human plasma by binding to a denatured gelatin column followed by elution with high concentrations of arginine. The eluted material was further purified by gel filtration.
Our Abpromise guarantee covers the use of ab299 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||Use at an assay dependent concentration.|
|IP||1/4000 - 1/8000.|
|WB||1/4000 - 1/8000.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|Sandwich ELISA||Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [HFN 7.1] to Fibronectin (ab80923). For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Mouse monoclonal [HFN 7.1] to Fibronectin - BSA and Azide free (ab80923) as Capture.|
IHC image of Fibronectin antibody staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab299, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.