Background Staining, Autofluorescence and Blocking Steps

Abstract

Background staining has increased the amount of gray hair on the heads of histochemists. In most cases, background staining is not caused by a single factor. Along with Fc receptors, frequent causes of background staining are endogenous enzyme activity, if you use peroxidase or alkaline phosphatase as enzyme markers on your secondary antibodies, and endogenous biotin when using a streptavidin or avidin reagents ( http://www.ihcworld.com/ ; http://www.protocol-online.org/prot/Immunology/ ). When fluorescent dyes are used in experiments, autofluorescence (or natural fluorescence) of some tissue components can cause background problems and complicate the use of fluorescence microscopy.