Basic Protein and Peptide Protocols

1994. Auflage 1994
ISBN: 978-0-89603-268-2
Verlag: Humana Press

Buch, Englisch, Band 32, 490 Seiten, Format (B × H): 156 mm x 234 mm, Gewicht: 1940 g

Reihe: Methods in Molecular Biology

The first volume in this Methods Molecular Biology series, Proteins (1984), concentrated on basic techniques for the analysis and purification of peptides and proteins. As the series developed, more specialized volumes on proteins were introduced, such as those on Immunochemical Protocols (vol. 10), Practical Protein Chro- tography (vol. 11), Analysis Glycoprotein Biomedicine (vol. 14), Protein-DNA Interactions (vol. 30), Biomembrane Protocols (vols. 19 and 27), Analyses and Methods (vol. 17), and Optical Spectroscopy, Microscopy, and Macroscopic Techniques (vol. 22). Further specialist volumes on peptides, monoclonal antibodies, immunoassays, ELISA, protein engineering, protein stability, mass spectrometry of proteins, automated sequence analysis, and protein NMR are currently in preparation. Since it is now a decade since the initial volume was published, it seems an especially appropriate moment to extensively reorganize, update, and revise the earlier volume. In an attempt to be more c- prehensive in our coverage, this current volume, Basic Protein and Peptide Protocols, is totally committed to basic analytical methods; a planned companion volume will later concentrate on preparative techniques. Those analytical techniques requiring expensive speci- ized instrumentation, such as NMR, mass spectrometry, X-ray cr- tallography, spectroscopy, and automated sequence analysis, are not described here, but in the appropriate specialized volumes listed above.

Zielgruppe

Research

Autoren/Hrsg.

Walker, John M

Fachgebiete

• Interdisziplinäres Wissenschaften Wissenschaften: Forschung und Information Forschungsmethodik, Wissenschaftliche Ausstattung
• Naturwissenschaften Biowissenschaften Proteinforschung
• Naturwissenschaften Biowissenschaften Biochemie (nichtmedizinisch)

Inhaltsverzeichnis

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 of Proteins.- 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.- 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 Protein 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-Performance 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.- 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 Bands.- 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 Immunosorbent Assay (ELISA).- Free Zone Capillary Electrophoresis.