This manual follows at a distance of 3 years the previous one entitled Mem brane Proteins, and, like its predecessor, it is the result of an International Advanced Course sponsored by FEBS, SKMB and SNG, which was held in Bern in September 1983. The experiments offered to the students in the course had to be largely up· dated or chosen from new areas of membrane research, because of the sub stantial and rapid development of the field. Using the protocols of the course, the participants (graduate students, postdoctoral fellows and also senior scientists), in most cases not at all ex pert in biomembrane research, were able to repeat all the experiments suc cessfully. Those few protocols which for some reason did not fulfill the role we expected were modified. These protocols have now been collected in this manual, which we are able to offer to a number of biology, biochemistry and biophysics laborato ries, hoping that the selected number of methods which have been success fully used during the Advanced Course may be useful to them. This manual is also intented for teachers of practical classes, who may use it as a text book and as source of selected references, collected not in the library, but in the laboratory, from the notebooks of the young researchers who have contributed so much to the success of the Course.

I. Analytical Separation Techniques

Electrophoretic Transfer of Chloroplast Membrane Proteins from SDS-Gels onto Nitrocellulose and Their Immunological Detection
Investigation of Amphiphilic Nature of Different Forms of Acetylcholinesterase from Torpedo Marmorata by Charge Shift Crossed Immunoelectrophoresis
Separation of Hydrophobic Membrane Proteins by Phase Partition: Characterization by Two-Dimensional Gel Electrophoresis and Silver-Staining
Quantitative Analysis of Plant Membrane Lipids by a Combined TLC-GLC Procedure
Localization of Phosphatidylglycerol in the Membrane of Acholeplasma laidlawii
II. Protein Structure and Interactions
Crystallization of Two Membrane Proteins: Bacteriorhodopsin and Photosynthetic Reaction Centres
Investigation by Crossed Immunoelectrophoresis of Membrane-Cytoskeleton Interactions in Human Erythrocyte Membranes
III. Labeling and Crosslinking
Labeling of the Hydrophobic Core of Membranes with 3-trifluoromethyl-3-(m-[125I] iodophenyl) diazirine: Measurement of the Time-Course of the Photolabeling Process
Selective Labeling of the ADP/ATP Translocator with Eosin-5-Maleimide
Site-Directed Hydrophobic Labeling of Membrane Proteins: NBD-Modification of Bacteriorhodopsin
IV. Reconstitution
Reconstitution of Na, K-ATPase
Import of Proteins by Isolated Mitochondria
Isolation and Functional Reconstitution of Rat Liver Cytochrome Oxidase
V. Spectral Techniques
Fluorescent Labeling of Band 3 Protein from Erythrocytes
Spin Labeling of Membranes and Membrane Proteins
Spectroscopic Measurements in the Subsecond to Second Time Domains: Use of an Optical Multichannel Analyzer for Spectral and Temporal Data Acquisition
Use of a Potential Sensitive Dye in Studies on Phospholipid and Sarcoplasmic Reticulum Vesicles
VI. Receptors
Beta-Adrenoceptor Regulation in Organs of Rats Chronically Treated with Reserpine or Isoproterenol. Radioligand-Receptor Binding Studies
Functional Changes in the Beta-Adrenoceptor Adenylate Cyclase System During Reticulocyte Maturation in vitro
Acetylcholine Receptor-Enriched Membrane Fragments from the Electric Organ of Torpedo Marmorata.