"Rainbow" western blotting.

ABSTRACT

The ''Rainbow western'' method permits detection of multiple antigens on a single protein blot. The procedure utilizes horseradish peroxidase (HRPase)-based detection with both a chemiluminescent and colorimetric substrate. In the ''Rainbow western'' procedure four different HRPase-colorimetric substrates that produce black, brown, red, and green colors are employed sequentially for detection and simultaneous display of four different antigens on the same blot. The Rainbow western methods have the potential to consolidate the work to analyze the expression levels of several proteins in studies of signaling pathways within biological samples. This technique could be particularly valuable for analysis of comigrating proteins, isoforms, and/or facilitating studies on phosphorylation, acetylation, and oligomerization of proteins tagged by the same epitope.