Manual Immunofluorescence Staining for Dystrophin and Key Dystrophin-Associated Proteins.

Abstract

The detection of dystrophin and the proteins of the dystrophin-associated protein complex (DAPC) by immunostaining allows for the detection of the presence and localization of these proteins in slide-mounted sections of skeletal muscle or heart tissue. This technique has been used historically for the diagnosis and characterization of muscle disorders including Duchenne muscular dystrophy and many of the limb girdle and congenital muscular dystrophies. The extensive usefulness of this technique has allowed the development of numerous antibodies that can be used in immunostaining assays. As an increasing number of research and translational science studies have recently focused on the restoration of dystrophin as a therapeutic strategy, high-quality immunostaining techniques and analysis remain sought-after scientific practices. This chapter describes useful procedures to perform the immunofluorescence staining for dystrophin and several other useful DAPC proteins, while also providing guidance based on our clinical diagnostic and translational science research experience from working in the muscular dystrophy field.