The choice of blocking agent depends on several factors, including the tissue type, the antibodies used, and the target antigen. Here are some considerations:
Tissue Type: Different tissues may have varying levels of endogenous proteins that could cause non-specific binding. For example, BSA is commonly used for general purposes, while normal serum might be preferred for more sensitive tissues. Antibodies: The species and type of antibodies used can influence the choice of blocking agent. For instance, if using mouse antibodies, normal mouse serum might be used to block Fc receptors. Target Antigen: The characteristics of the target antigen, such as its abundance and location, can also dictate the blocking strategy. Highly abundant antigens may require more stringent blocking to differentiate between specific and non-specific signals.
