Western Blot Analysis With Chemiluminescent Detection of Proteins Conjugated to Fatty Acid Ethyl Esters

Tsai, Hsing-Chen, and Elizabeth Roberts-Kirchhoff

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were used for the analysis and definition of proteins conjugated with and without fatty acid ethyl esters. SDS-PAGE was used to separate proteins based on size with visualization by staining with Coomassie blue dye. Western blotting was used to transfer the proteins from a gel to a nitrocellulose membrane followed by the characterization of the proteins with antibodies and chemiluminescent detection. Conditions for the SDS-PAGE and Western blot analysis were determined for the limited tryptic digestion of bovine serum albumin, bovine serum albumin conjugated with ethanol-esterified 13-hydroxyoctadeca-9,11-dienoic acid, horseradish peroxidase and horseradish peroxidase conjugated with ethanol-esterified 13-hydroxyoctadeca-9,11- dienoic acid.