The choice of blocking agent depends on several factors, including the type of tissue, the nature of the antigen, and the specific antibodies used. Here are some guidelines:
Type of Tissue: For most tissues, BSA or normal serum works well. However, for more complex tissues, such as brain or liver, a combination of blocking agents may be necessary. Nature of Antigen: For antigens that are very abundant, a stronger blocking agent like casein may be required. For less abundant antigens, BSA or normal serum may suffice. Antibodies Used: The species of the primary and secondary antibodies can influence the choice of blocking agent. Using normal serum from the same species as the secondary antibody can be particularly effective in reducing non-specific binding.
