Biological and Biomedical Applications of Isoelectric Focusing

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1 Isoelectric Focusing of Human Saliva, Cerebrospinal Fluid, and Urine.- I. Introduction.- II. Isoelectric Focusing of Saliva.- A. Gel Rods and Thin Layer Gels.- B. Liquid Columns.- C. ?-Amylases.- D. Immunoglobulins.- E. Blood Group Specific Substances.- F. Vitamin B12 Proteins Binding Proteins (Cobalophilins).- G. Other Glycoproteins.- H. Lymozyme.- I. Specific Salivary Proteins.- J. Dental Plaque.- K. Other Proteins.- III. Isoelectric Focusing of Cerebrospinal Fluid.- IV. Isoclectdc Focusing of Urine.- V. Conclusions.- VI.References.- 2 The Use of Gel in the Analysis of Hemoglobins.- I. Introduction.- II. Practical Applications.- A. Normal Human Hemoglobins.- B. Clinical Studies.- C. Identification of Hemoglobin Variants.- D. Subunit Identification.- E. Identification of Animal Hemoglobins.- III. Experimental Applications.- A. Preparation of Purified Hemoglobins.- B. Conformational Isomerization.- C. Asymmetrical Hybrids.- D. Valence Hybrids.- E. Chemical Modification.- IV. References.- 3 Isoelectric Focusing of Allergens.- I. Introduction.- II. Methods.- A. Isoelectric Focusing Method Used for the Characterization of Allergens.- B. Paper Disk Radio- Allergo-Sorbent Test (Paper Disk RAST) Used for the Detection of Allergens.- C. The Preparation of Agents for the Detection of Allergens.- D. The Effect of Freezing Allergens.- III. Isoelectric Focusing of Allergens.- A. Effect of Polyacrylamide Gel Concentration.- B. Effect of Different Times of Electrophoresis.- C. Reproducibility of Method.- D. Correlation of I soelectrophoretically Separated Allergen Components with Protein Patterns.- IV. Applications.- A. Allergen Dose-Response Relationship.- B. Variability in pl Component Spectrum of Allergens Determined with Several Allergen Sensitive Sera.- C. Differences in pl Component Spectra of Allergen Extracts Prepared by Different Producers.- D. Changes in pl Component Spectra of Allergen Extracts Resulting from Storage.- E. Changes in Dose-Response Relationships and in Isoelectric Focusing Patterns of Allergen Extracts Treated with Enzymes.- F. Detection of Allergens by Enzyme-Linked Allergo-Sorbent Test (ELAST).- G. Isoelectric Focusing Patterns of Some Purified Allergens.- V. General Comments.- VI. References.- 4 Isoelectric Focusing of Muscle Proteins.- I. Introduction.- II. Technical Considerations.- A. Preparative Procedures.- B. Analytical Procedures.- III. Specific Applications.- A. Preparations and Isolations of Muscle Proteins.- B. Analyses of Muscle Proteins.- C. Applications of Special Interest.- IV. Conclusions and Predictions.- V. References.- 5 Studies of the Structure and Synthesis of Immunoglobulins by Isoelectric Focusing.- I. Introduction.- II. Methods of Isoelectric Focusing of Immunoglobulins.- A. Liquid Density Gradient Stabilized Columns.- B. Acrylamide Gel Stabilized Methods.- C. Granular Gel Stabilized Methods.- D. The Use of Urea for Solubilization.- E. Isoelectric Focusing of Immunoglobulin Chains and Fragments.- III. Isoelectric Focusing of M-Proteins.- A. Focusing Patterns of Monoclonal Immunoglobulins.- B. The Nature of Immunoglobulin Microheterogeneity.- IV. Studies of Antibody Structure and Heterogeneity by Isoelectric Focusing.- A. Antihapten Antibodies.- B. Anticarbohydrate Antibodies.- C. Antiprotein Antibodies.- V. Studies of Antibody Synthesis.- A. In Vivo Cloning.- B. In Vitro Cloning (Monofocal Antibodies).- C. Estimates of Antibody Diversity in a Single Animal.- D. Studies of Antibody Production in Neonatal Animals.- E. Clonal Dominance.- F. Gene Linkage Studies.- VI. References.- 6 The Use of Isoelectric Focusing in the Separation and Characterization of Nuclear Nonhistone Proteins.- I. Introduction.- II. General Properties of NHP.- III. Isoelectric Focusing of NHP.- A. Separation of NHP from Histones.- B. Analyses of NHP Using Isoelectric Focusing.- C. Preparation of NHP Using lsoelectnc Focusing.- IV. Artifacts and the Heterogeneity of NHP.- A. General.- B. Nucleic Acids.- C. Modifications of NHP.- D. Effect of Denaturing Agents.- E. Binding of Ampholines.- F. Effects of Extremes of pH.- G. Effects of Proteolytic Activity.- V. Conclusion.- VI. References.- 7 Isoelectric Focusing of Membrane Components.- I. Introduction.- II. Methods.- A. Solubilization of Membrane Proteins.- B. Focusing Procedure.- C. Two-Dimensional Techniques.- III. Biological Applications.- A. Erythrocyte Membranes.- B. Plasma Membranes from Lymphoid Cells.- C. Bacterial Membranes.- D. Chloroplast Membranes.- E. Membrane Lipids in Isoelectric Focusing.- IV. References.- 8 Isoelectric Focusing of Seed Proteins.- I. Introduction.- II. Food Proteins.- A. Cereals.- B. Oilseeds.- C. Other Plant Proteins.- III. Enzymes.- A. Carbohydrases.- B. Proteases and Peptidases.- C. Lipoxygenases.- D. Peroxidases.- E. Esterases.- F. Urease.- G. Myrosinase.- IV. Protease Inhibitors.- A. Soybean.- B. Groundnut.- C. Barley.- D. Potato.- V. Lectins.- A. Nutritional Effects.- B. Isolectins.- C. Aggregation.- D. Sugar Binding.- E. Subunit Structure.- VI. Genetic Studies.- A. Location of Genes Controlling Protein Synthesis.- B. Phylogenetic Relationships.- C. Variety Identification.- D. Food Analysis.- VII. Conclusion.- VIII. References.- 9 Isoelectric Focusing of Microbial Proteins.- I. Introduction.- II. Materials.- A. Ampholytes.- B. Anticonvection Media.- C. The Sample.- III. Methods.- IV. Isoelectric Focusing of Microbial Proteins.- A. Viral Proteins.- B. Protozoan Proteins.- C. Fungal Proteins.- D. Bacterial Proteins.- V. Conclusions.- VI. References.- 10 Isoelectric Focusing of Serum Proteins.- I. Introduction.- II. Gel Isoelectric Focusing Alone.- A. Gel Isoelectric Focusing in One Dimension.- B. Gel Rod Electrofocusing of Whole Serum and Plasma Proteins.- III. Gel Electrofocusing Followed by Electrophoresis.- A. Identification of Human Serum Protein Zones.- B. Animal Sera.- IV Individual Types of Protein in Serum or Plasma.- A. Albumin.- B. ?1-Antitrypsin.- C. Coagulation and Fibrinolytic Factors.- D. ?1-Fetoprotein.- E. Glycoproteins.- F. Haptoglobins.- G. Hemopexin.- H. Immune-Response Proteins.- I. The Lipoproteins.- J. ?2-Macroglobulin.- K. Transport Proteins.- L. Hormones and Related Proteins.- M. Miscellaneous Observations.- V. Clinical Applications.- A. Australia Antigen.- B. Cystic Fibrosis.- C. Gammopathies.- D. Lipoprotein Disorders.- E. Miscellaneous.- VI. References.