New Protein Techniques: Methods in Molecular Biology, cartea 3

Prevention of Unwanted Proteolysis.- The Bradford Method for Protein Quantitation.- Amino Acid Analysis by Precolumn Derivatization.- Identification of N-Terminal Amino Acids by High-Performance Liquid Chromatography.- Enzymatic Methods for Cleaving Proteins.- Chemical Cleavage of Proteins.- Chemical Modification of Proteins.- The Design, Preparation, and Use of Immunopurification Reagents.- Dye-Ligand Chromatography.- Aminohexyl-Sepharose Affinity Chromatography.- Purification of DNA-Dependent RNA Polymerase from Eubacteria.- Direct Immunoprecipitation of Protein.- Detection of Proteins in Polyacrylamide Gels Using an Ultrasensitive Silver Staining Technique.- Chromatofocusing.- Hybrid Isoelectric Focusing Using Mixed Synthetic-Carrier Ampholyte-Immobilized pH Gradient Gels.- Two-Dimensional Electrophoresis Using Immobilized pH Gradients in the First Dimension.- Two-Dimensional Polyacrylamide Gel Electrophoresis Using Flat-Bed Isoelectric Focusing in the First Dimension.- Preparative Aspects of Immobilized pH Gradients.- Isoelectric Focusing Under Denaturing Conditions.- Computer Analysis of 2-D Electrophoresis Gels.- Two-Dimensional (Crossed) Immunoelectrophoresis.- Peptide Synthesis.- Synthesis of a Series of Analogous Peptides Using T-Bags.- Production of Antisera to Synthetic Peptides.- Production of Antibodies Using Proteins in Gel Bands.- Purification of Immunoglobulins Using Protein A-Sepharose.- Antibody-Enzyme Conjugate Formation.- Vacuum Blotting.- Blotting with Plate Electrodes.- Use of Dried Milk for Immunoblotting.- Immunodetection of Proteins on “Western” Blots Using 125I-Labeled Protein A.- Detection of Protein Blots Using the Avidin-Biotin System.- Detection of Protein Blots Using Enzyme-Linked Second Antibodies or Protein A.- Colloidal Gold for theDetection of Proteins on Blots and Immunoblots.- Enzyme Immobilization by Adsorption.- Enzyme Immobilization by Entrapment.- Enzyme Immobilization by Covalent Bonding.- Cell Immobilization.