Analysis of Proteins

Abstract

Abstract This overview discusses a broad array of aspects involved in protein purification, including historical background, determining the purpose for purifying a particular protein, and methods and recommendations for specific purification procedures. Strategies are discussed based on the classification of the protein according to shape, oligomeric structure, function, as well as location and associations within the cell. Specifically, critical issues are detailed for extracellular, cytoplasmic, and membrane‐associated proteins, as well as for insoluble proteins. Strategies for purification of both soluble and insoluble (inclusion bodies) recombinantly expressed proteins are also provided. Protein purification flow charts are presented to give a broad outline of the methods used for different types of proteins. Also covered are methods of analysis and detection, and techniques for characterizing a purified protein.