ABSTRACT
Contents 20.1 Introduction ......................................................................................................................... 244 20.2 Starting Materia1 .................................................................................................................. 245
20.2.1 Purification of Thylakoid Membranes .................................................................. 245 20.2.1.1 Method for Higher Plants .................................................................... 245 20.2.1.2 Method for Most Unicellular Algae .................................................... 246 20.2.1.3 Method for Diatoms ............................................................................. 247
20.3 Detergent Solubilization ...................................................................................................... 248 20.3.1 Types of Detergents ............................................................................................... 248 20.3.2 Detergent Solubilization of Thy1akoid Membranes ............................................. 248
20.4 Resolution of Pigment-Protein Complexes ........................................................................ 249 20.4.1 Mild "Green Gel" LDS-PAGE .............................................................................. 249 20.4.2 Sucrose Density Gradient Ultracentrifugation ...................................................... 250 20.4.3 Column Chromatography ...................................................................................... 252
20.5 Biochemical Characterization of Pigment-Protein Complexes ......................................... 252 20.5.1 Determination of Polypeptide Composition ......................................................... 252
20.5.1.1 Composition of Gels and Running Buffers ......................................... 252 20.5.1.2 Sample Preparation .............................................................................. 253
20.5.1.3
20.5.1.4
20.5.1.2.1 Gel Slices from "Green Gels" ......................................... 253 20.5.1.2.2 Sucrose Density Gradient Fractions ................................ 253 20.5.1.2.3 Preparation of Standards ................................................. 253 Visualization of Polypeptides .............................................................. 254 20.5.1.3.1 High-Sensitivity Coomassie Blue Stain .......................... 254 20.5.1.3.2 Silver-Staining Method .................................................... 254 Immunochemical Identification of Proteins ........................................ 254
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20.5.2DeterminationofPigmentComposition...............................................................255 20.5.2.1ExtractionofPigmentsUsingAcetone...............................................255
