Basic Protein and Peptide Protocols

Abstract

The Lowry Method for Protein Quantitation. The Bicinchoninic Acid (BCA) Assay for Protein Quantitation. The Bradford Method for Protein Quantitation. Nondenaturing Polyacrylamide Gel Electrophoresis of Proteins. SDS Polyacrylamide Gel Electrophoresis of Proteins. Gradient SDS Polyacrylamide Gel Electrophoresis. Acetic Acid-Urea Polyacrylamide Gel Electrophoresis of Proteins. Electrophoresis of Peptides. Isoelectric Focusing of Proteins in Ultra-Thin Polyacrylamide Gels. The In Vivo Isotopic Labeling of Proteins for Polyacrylamide Gel Electrophoresis. Two-Dimensional Polyacrylamide Gel Electrophoresis of Proteins. Two-Dimensional Polyacrylamide Gel Electrophoresis Using Immobilized pH Gradients in the First Dimension. Two-Dimensional Polyacrylamide Gel Electrophoresis Using Flat-Bed Isoelectric Focusing in the First Dimension. Quantification of Proteins on Polyacrylamide Gels (Nonradioactive). Detection of Proteins in Polyacrylamide Gels Using an Ultrasensitive Silver Staining Technique. Identification of Glycoproteins on Nitrocellulose Membranes and Gels. The Release of Oligosaccharides from Glycoproteins. Structural Profiling of Oligosaccharides of Glycoproteins. Drying Gels. Fluorography of Polyacrylamide Gels Containing Tritium. The Electrophoretic Elution of Proteins from Polyacrylamide Gels. Preparation of Proteins from Gels for Protein Microsequencing. Comparison of Protein Primary Structures: Peptide Mapping. Protein Blotting. Detection of Polypeptides on Immunoblots Using Secondary Antibodies or Protein A. Detection of Proteins on Blots Using the Avidin-Biotin System. Detection of Proteins on Blots Using Chemiluminescent Systems. The Detection of Proteins on Blots Using Gold or Immunogold. Amino Acid Analysis. Molecular-Weight Estimation for Native Proteins Using Size-Exclusion High-Pressure Liquid Chromatography. Peptide Purification by Reverse-Phase HPLC. Enzymatic Methods for Cleaving Proteins. Chemical Cleavage of Proteins. Chemical Modification of Proteins. The Dansyl Method for Identifying N-Terminal Amino Acids. The Dansyl-Edman Method for Peptide Sequencing. C-Terminal Sequencing of Peptides: The Thiocyanate Degradation Method. Analysis of Cysteine Residues and Disulfide Bonds. Preparation and Characterization of Monoclonal Antibodies to Proteins and Other Cellular Components. The Raising of a Polyclonal Antiserum to a Protein. Production of Antisera to Synthetic Peptides. Production of Antibodies Using Proteins in Gel Bonds. Preparation of Purified Immunoglobulin G (IgG). Enzyme and Biotin Labeling of Antibody. Labeling of Peptides and Proteins by Radioiodination. Radioimmunoassay of Peptides and Proteins. Enzyme-Linked Immunosorbant Assay (ELISA). Free Zone Capillary Electrophoresis. Index.