ABSTRACT
This chapter presents Immunofluorescence methods and techniques for visualizing the subcellular location of gap junction proteins and their associated proteins. It serves as a guide for researchers interested in the morphological analysis of gap junctions. The chapter describes protocols for localizing connexin in cells on coverslips by immunofluorescence techniques with quantum dots (Qdots). It includes protocols for Qdot immunoelectron microscopy of cells on coverslips. The selection of the appropriate fixative is a key to good immunofluorescence results. A good fixative should prevent the loss of cellular structures without destroying the antigen-binding sites needed to bind the primary antibody. Secondary antibodies are available from a number of commercial sources. A secondary antibody that is compatible with the primary antibody must be used. In addition to membrane staining, actin is sometimes used to distinguish the cell outlines and to study the relationships of connexin to the cytoskeleton.
