Methods in molecular biology. 19, Biomembrane protocols

Preface v Contributors ix CH. 1. The Identification of Subcellular Fractions from Mammalian Cells, John M. Graham 1 CH. 2. The Isolation of Nuclei and Nuclear Membranes from Rat Liver, John M. Graham 19 CH. 3. Isolation of Mitochondria, Mitochondrial Membranes, Lysosomes, Peroxisomes, and Golgi Membranes from Rat Liver, John M. Graham 29 CH. 4. Continuous-Flow Electrophoresis: Application to the Isolation of Lysosomes and Endosomes, John M. Graham 41 CH. 5. Fractionation of a Microsomal Fraction from Rat Liver or Cultured Cells, John M. Graham 57 CH. 6. Isolation of Plasma Membrane Sheets and Plasma Membrane Domains from Rat Liver, Laura Scott, Michael J. Schell, and Ann L. Hubbard 59 CH. 7. The Analysis of Receptor-Mediated Endocytosis by Centrifugation in Isoosmotic Gradients, Terry C. Ford 71 CH. 8. The Isolation of Clathrin-Coated Vesicles and Purification of Their Protein Components, Antony P. Jackson 83 CH. 9. Isolation of Membranes from Tissue Culture Cells, John M. Graham 97 CH. 10. The Isolation of Membranes from Bacteria, Robert K. Poole 109 CH. 11. Isolation and Purification of Functionally Intact Chloroplasts from Leaf Tissue and Leaf Tissue Protoplasts, David G. Whitehouse and Anthony L. Moore 123 CH. 12. Isolation and Purification of Functionally Intact Mitochondria from Plant Cells, Anthony L. Moore, Anne-Catherine Fricaud, Andrew J. Walters, and David G. Whitehouse 133 Vll via Contents CH. 13. Isolation of Cholinergic-Specific Synaptosomes by Immunoadsorption, J. Paul Luzio and Peter J. Richardson 141 CH. 14. Separation and Analysis of Phospholipids by Thin Layer Chromatography, Ian J. Cartwright 153 CH. 15. Analysis of Phospholipids by High Performance Liquid Chromatography, Hafez A. Ahmed 169 CH. 16. Measurement of Cholesterol in Membranes, Hafez A. Ahmed 179 CH. 17. Analysis of Fatty Acids by Gas Liquid Chromatography, Ian J. Cartwright 183 CH. 18. Chemical Assays for Proteins, David J. Winterbourne 797 CH. 19. Separation and Analysis of Membrane Proteins by SDS-Polyacrylamide Gel Electrophoresis, Michael J. Dunn and Samantha J. Bradd 203 CH. 20. Analysis of Membrane Proteins by Western Blotting and Enhanced Chemiluminescence, Samantha J. Bradd and Michael J. Dunn 277 CH. 21. Two-Dimensional Polyacrylamide Gel Electrophoresis of Membrane Proteins, Joe Corbett and Michael J. Dunn 279 CH. 22. Isolation of a Membrane-Bound Enzyme, 5'-Nucleotidase, / . Paul Luzio and Elaine M. Bailyes 229 CH. 23. The Isolation of Membrane Proteoglycans, Malcolm Lyon 243 CH. 24. Oligosaccharide Mapping and Sequence Analysis of Glycosaminoglycans, Jeremy E. Turnbull 253 CH. 25. Analysis of Sugar Sequences in Glycoproteins by Glycosidase Digestion and Gel Filtration, Anthony P. Corfield 269 CH. 26. The Extraction and Analysis of Glycosphingolipids, Norman A. Gregson 287 Appendix: 1. Density Gradient Media 303 Appendix: 2. Balanced Salt Solutions 304 Index 305