Electrophoresis of Proteins in Polyacrylamide and Starch Gels

Chapter 1. Applicability of Starch and Acrylamide Zone Electrophoresis Introduction Analytical Applications Preparative Applications Properties of Starch and Acrylamide Gels Movement of Fluid Through Gels Adsorption Effects - Band Shapes Mechanical Properties Transparency Gels Suitable for Separation on the Basis of Charge Difference Alone Buffers Suitable for Electrophoresis in Gels The Same Buffer in the Gel and in the Electrode Vessels Different Buffers in the Gel and in the Electrode Chambers Exchange of Buffer after Gel Has Been Set Gels Containing Solubilisers Urea Sodium Dodecyl Sulphate Phenol and Acetic Acid Evolution of Heat and Permissible Thickness of Gels Chapter 2. Acrylamide Gel Choice of Gel Choice of Buffer Solutions Acrylamide Gel Rendered Soluble after Use Catalysts Proportion of Bisacrylamide to Total Acrylamide Chapter 3. Apparatus and Techniques Choice of Apparatus Apparatus Suitable for Gels Which are to be Stained Optimum Conditions for Electrophoresis in Vertical Glass Tubes; 'Disc Electrophoresis' Preparation of Gel (An Example) Electrode Buffer Sample Solution Addition of Sample Solution Time and Current Required for Electrophoresis Removal of Gel Cylinders from Glass Tubes Use of Longer Gels Suitable for Disintegration by Extrusion Staining and Destaining Vertical Slab Between Cooling Plates Preparation of Gel and Leakage Check Entry of Proteins into Gels by Preliminary Passage of Current Sample Insertion Possible Reasons for Distortion of Bands Voltage, Current and Time of Electrophoresis Two-Dimensional Techniques Staining Methods Spectrophotometry of Acrylamide Gels Drying and Storage of Stained Acrylamide Gels Autoradiographs Estimation of Radioactivity in Acrylamide Gel Chapter 4. Methods Allowing Recovery of Separated Materials {Preparative Methods) Advantages and Disadvantages of Acrylamide Gels Insolubility Flexibility and Resistance to Cutting Resistance to Disintegration Stickiness and Adherence to Glass Swelling Formation of Gels with a Flat Upper Surface Methods of Employment and Types of Apparatus Elution from Sections of Gel Before or after Disintegration Vertical Columns with End Elution Shape And Degree of Separation of Peaks Obtained by Continuous Elution Apparatus of Gordon and Louis Apparatus of Hjerten et al Apparatus of Duesberg and Rueckert Apparatus of BrownstoneChapter 5. Starch Gel Introduction Pretreatment of Granular Starch in Acid Acetone Choice of Appropriate Starch Gel Pore Size Buffer Solution in the Gel Effect of Urea When Present in Starch Gel Buffer Systems for Particular Proteins (Starch Gel) Preparation of Starch Gel Choice of Apparatus Horizontal Apparatus (Normal Size) Horizontal Apparatus (Micro Scale) Vertical Apparatus Separations in Two Dimensions Two-Dimensional Apparatus Treatment of Starch Gel after Electrophoresis Slicing Staining Densitometry Autoradiographs Special Adaptations for Recovery of Materials after Separation in Starch Gel Removal of Soluble Starch Acknowledgments AppendixReferences Subject Index