Luminescent immunodetection of western-blotted proteins from coomassie-stained polyacrylamide gel

Abstract

Immunodetection with horseradish peroxidase-linked antibodies on Coomassie-stained nitrocellulose blots can be performed efficiently and rapidly with the peroxidase substrate luminol. The luminescence produced is detected with radioautographic film. This procedure allows a direct identification of immunodetected bands of stained nitrocellulose sheets without using radiolabeled secondary antibodies. Because of its convenience and sensitivity, this method could be particularly suitable for purification of immunodetected proteins.