Blocking is a crucial step in fluorescence microscopy to prevent nonspecific binding of antibodies and reduce the occurrence of artifacts.
Protocol
Typically, either a 3% solution of Bovine Serum Albumin (BSA), Normal Goat Serum (NGS), or Normal Donkey Serum (NDS) is used for this purpose. This blocking solution is applied to the cells for an incubation period of approximately 20 minutes. By doing this, potential sites where antibodies could bind nonspecifically are ‘blocked’, allowing for the specific binding of the antibodies to their targets in subsequent steps of the protocol. This increases the signal-to-noise ratio and produces cleaner, more interpretable results. However, many procedures are often modified to suit a particular purpose.
