Strategies for Handling Polypeptides on a Microscale -- SDS Polyacrylamide Gel Electrophoresis for N-Terminal Protein Sequencing -- Two-Dimensional Polyacrylamide Gel Electrophoresis for the Separation of Proteins for Chemical Characterization -- Electroblotting of Proteins from Polyacrylamide Gels for Chemical Characterization -- Enzymatic Cleavage of Polypeptides -- Chemical Cleavage of Polypeptides -- Preparation of Glycopeptides -- Selective Isolation of the Carboxy Terminal Peptide from a Protein -- Selective Isolation of the Amino-Terminal Peptide from ?-Amino Blocked Protein -- Analytical and Micropreparative Capillary Electrophoresis of Peptides -- Reverse-Phase HPLC Purification of Peptides from Natural Sources for Structural Analysis -- High-Performance Liquid Chromatography On-Line Derivative Spectroscopy for the Characterization of Peptides with Aromatic Amino Acid Residues -- Hydrolysis of Samples for Amino Acid Analysis -- Amino Acid Analysis -- Postcolumn Amino Acid Analysis -- Fast-Atom Bombardment and Electrospray Mass Spectrometry of Peptides, Proteins, and Glycoproteins -- Peptide Mass Fingerprinting Using MALDI-TOF Mass Spectrometry -- N-Terminal Protein Ladder Sequencing -- The Dansyl-Edman Method for Manual Peptide Sequencing -- The Dansyl Method for Identifying N-Terminal Amino Acids -- Identification of PTH-Amino Acids by High-Performance Liquid Chromatography -- Protein Sequencer Maintenance and Troubleshooting -- Identifying the Carboxy Terminal Amino Acid of a Protein -- Enzymatic and Chemical Methods for Manual C-Terminal Peptide Sequencing -- Automated Methods for C-Terminal Protein Sequencing -- Chemical Modification of Proteins for Sequence Analysis -- On-Membrane Deblocking of Proteins -- Analysis of Sites of Protein Phosphorylation -- Identification of Sites of Glycosylation -- Quantification and Location of Disulfide Bonds in Proteins.

Bryan Smith draws together in Protein Sequencing Protocols a timely collection of current and state-of-the-art techniques for protein and peptide preparation and sequencing-all described by hands-on masters of the procedures. These well-tested methods convey valuable lessons and insights into using automated sequencers and/or mass spectrometers, preparing proteins and peptides on a microgram and submicrogram scale, mapping peptides, analyzing amino acid by pre- and post-column derivatization methods, and sequencing N- and C-terminal peptides by classical and newly developed techniques. They also deal with such troublesome problems as identifying sites of modification and handling very small samples. Protein Sequencing Protocols offers self-contained, step-by-step recipes that ensure easily reproducible results for those already doing sequencing, and for those who are still learning the methods, as well as for those already in one area but wishing to learn associated techniques. Its multiplicity of proven protocols will guide the challenged experimenter to successful results.