Cellular proteins that interact with p53 play a major role in both positive and negative regulation of this tumor suppressor and can fine-tune its response to specific cellular stresses. As a consequence, p53 biology will not be complete until these interacting proteins are fully characterized. This chapter outlines two methods for identifying and characterizing p53-binding proteins: (i) glutathione-S-transferase (GST) protein—protein interaction assay; and (ii) co-immunoprecipitation (co-IP) assay. These two methods are ideal for any laboratory to perform; the assays are short in duration, do not require specialized expertise to establish in the laboratory, give reliable and reproducible data, and are cost-efficient, because few reagents are needed. This chapter gives a basic description of these two techniques and provides tips that are not found in other protocol manuals on how to achieve the best results.
