Strategies for Protein Purification and Characterization: A Laboratory Course Manual

Investigators who have identified and cloned a gene of interest often want to isolate and characterize the protein product, yet the methods required are notoriously difficult for the inexperienced. This manual has been adapted from a course held at the Cold Spring Harbour Laboratory in New York, to teach scientists how to execute the major protein techniques by applying them to four distinct, representative types of molecule: a regulatory protein, a DNA-binding protein, a recombinant protein, and a membrane-bound receptor. This manual covers a variety of bulk fractionation, electrophoretic, and chromatographic techniques. Step-by-step protocols are accompanied by troubleshooting advice and guidance on generalizing the techniques for other classes and types of protein. The emphasis throughout is on strategies for purification and characterization rather than automated instrumental analysis. The book should be of use to specialists in genetics, microbiology, neuroscience, and cell biology, who wish to develop expertise in working with proteins.