Biomembrane Protocols, Volume 1: Isolation and Analysis

About this book

Expert practitioners combine the traditional basic techniques of membrane isolation and analysis with modern advances in molecular biology, immunology, and clinical sciences to devise analytical techniques that are successful for expert and novice alike. The techniques are used in the isolation and compositional analysis of membranes and address animal, plant, and microbial cells. The topics covered in this first volume include the isolation of subcellular fractions, centrifugation, the extraction and quantitation of lipids, the separation and analysis of proteins, and the isolation and characterization of glycosylated molecules, glycoproteins, proteoglycans, and glycolipids.

Contents

The Identification of Subcellular Fractions from Mammalian Cells. The Isolation of Nuclei and Nuclear Membranes from Rat Liver. Isolation of Mitochondria Mitochondrial Membranes Lysosomes Peroxisomes and Golgi Membranes from Rat Liver. Continuous-Flow Electrophoresis: Application to the Isolation of Lysosomes and Endosomes. Fractionation of a Microsomal Fraction from Rat Liver or Cultured Cells. Isolation of Plasma Membrane Sheets and Plasma Membrane Domains from Rat Liver. The Analysis of Receptor-Mediated Endocytosis by Centrifugation in Isoosmotic Gradients. The Isolation of Clathrin-Coated Vesicles and Purification of Their Protein Components. Isolation of Membranes from Tissue Culture Cells. The Isolation of Membranes from Bacteria. Isolation and Purification of Functionally Intact Chloroplasts from Leaf Tissue and Leaf Tissue Protoplasts. Isolation and Purification of Functionally Intact Mitochondria from Plant Cells. Isolation of Cholinergic-Specific Synaptosomes by Immunoadsorption. Separation and Analysis of Phospholipids by Thin Layer Chromatography. Analysis of Phospholipids by High Performance Liquid Chromatography. Measurement of Cholesterol in Membranes. Analysis of Fatty Acids by Gas Liquid Chromatography. Chemical Assays for Proteins. Separation and Analysis of Membrane Proteins by SDS-Polyacrylamide Gel Electrophoresis. Analysis of Membrane Proteins by Western Blotting and Enhanced Chemiluminescence. Two-Dimensional Polyacrylamide Gel Electrophoresis of Membrane Proteins. Isolation of a Membrane-Bound Enzyme 5'-Nucleotidase. The Isolation of Membrane Proteoglycans. Oligosaccharide Mapping and Sequence Analysis of Glycosaminoglycans. Analysis of Sugar Sequences in Glycoproteins by Glycosidase Digestion and Gel Filtration. The Extraction and Analysis of Glycosphingolipids. Index.

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