Membrane Biochemistry: A Laboratory Manual on Transport and BioenergeticsE. Carafoli, G. Semenza Springer Science & Business Media, 2012/12/06 - 178 ページ This manual collects in the form of laboratory protocols a series of experiments in the field of Membrane Transport and Membrane Bioenergetics. It represents the experience accumulated during four advanced courses held at the Depart ment of Biochemistry of the Swiss Federal Institute of Technology on behalf of Federation of European Biochemical Societies (FEBS) in the years 1975 through 1978. The idea of collecting the experiments into a laboratory manual developed as a response to a demand from the students who took part in the courses. Further motivation came with the fmding that, in planning the laboratory sessions, the teaching staff had no organized, modern source of information in the literature. The experiments presented cover most areas of importance in the subject mat ter. Their presentation has been continuously modified in the course of the four years during which the manual took shape, to accommodate to experience and various suggestions. In their present form, all of the experiments described have been repeatedly practiced to optimize their execution. Efforts have been made to combine in the manual classical experiments, and techniques which require relatively unsophisticated instrumentation and can therefore be carried out in most laboratories, with more modern experiments and relatively newer technol ogies. In its present form, the manual should therefore provide a usefui tool in the hands of researchers and laboratory teachers at different levels of sophisti cation and instrumentation. |
目次
1 | |
Transport of Sugars in Bacteria | 25 |
Net Nat and K+ Movements in Human Red Blood Cells After | 36 |
Calcium Transport in Sarcoplasmic Reticulum Vesicles Isolated | 51 |
Preparation and Assay of Animal Mitochondria and Submito | 62 |
Measurement of Cytochrome Kinetics in Rat Liver Mitochondria | 77 |
Proton Translocation Catalyzed by Mitochondrial Cytochrome | 82 |
Anion Transport in Mitochondria | 98 |
Redox Intermediates Between O₂ and H₂O 2 | 113 |
Oxygen Evolution and Uptake as a Measure of the LightInduced | 144 |
Characterization of Ionophores Using Artificial Lipid | 164 |
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acid added addition aliquots anion Antimycin aqueous assay ATPase bacteria bacteriorhodopsin Bchl Biochem Biochemistry Biochim Biophys Acta Biol Chem brane brush border buffer Ca² Ca2+ concentration Ca2+ transport CaCl calcium cells centrifuge chloroplasts chromatophores cocktail containing cuvette cytochrome oxidase D-glucose D-glucose transport determined dilution electron transport enzyme ethanol evol experiment EXPERIMENTAL PROCEDURES filter filtration final concentration flux glucose gradient H₂O HEPES incubation medium inhibition inhibitor intestinal INTRODUCTION AND AIMS ionophore ions kinetic lipid mannitol measured melibiose membrane potential mg/ml Mg2+ Millipore mitochondria murexide NaCl O₂ oligomycin oxidized oxygen electrode permeability permease phlorizin binding phosphate phosphorylation photophosphorylation photosynthetic pipette preincubated Preparation protein proton pump reaction mixture redox resealed respiratory rotenone sample sarcoplasmic reticulum spectrophotometer stock solution substrate succinate sucrose supernatant suspension translocation Tris-HCl tubes uncoupler uptake valinomycin vesicles vials volume wavelength Wikström