New Approaches to Quantitative Western Blotting

Fluorescent detection in Western blotting offers high sensitivity, broad dynamic range and stability of signals. This makes it highly suitable for quantitative Western blotting. Here we show how fluorescent Western blotting can be used for simultaneously detection of up to three different proteins on the same blot at the same time and for detection of proteins of the same molecular weight without stripping and reprobing. We also demonstrate how fluorescent Western blotting with 3 layer probing can be used to increase sensitivity and thereby enables detection of very low abundant proteins. Finally we demonstrate that it is possible to compare the total protein amount to the target protein by using Deep Purple™ protein staining prior to ECL™ Plex™ Western blotting.