Fractionation of Nucleic Acids on Isoelectric Focusing

Kapitel in diesem Buch

1. Frontmatter I
2. Preface V
3. Contents VII
4. Contributors XII
5. Part I. Isoelectric Focusing
6. 1. Theoretical and General Aspects
7. Reminiscences about the Genesis of Isoelectric Focusing and Generalization of the Idea 3
8. Stable pH Gradients - A Key Problem in Isoelectric Focusing 35
9. Electrofocusing in Buffers: Formation of Natural pH Gradients, Flexibility, Gradient Stability, Relation to Isotachophoresis and Preparative Potential 51
10. Comparison of pH Gradients Used for Isoelectric Focusing 59
11. On the Temperature Dependence of Isoelectric Points of Proteins with Special Reference to Isoelectric Focusing 71
12. 2. Carrier Ampholytes
13. Carrier Ampholyte Distribution 87
14. Characterization of Carrier Ampholytes by New Visualization Reactions and Chromatographic Methods 97
15. Synthesis of Carrier Ampholytes for Isoelectric Focusing Containing Sulfonic and Phosphonic Acid Groups Covering a Wide pH Range 111
16. 3. Analytical Methodology
17. Staining of Proteins in Polyacrylamide Gels 123
18. Isoelectric Focusing of Complex Protein Mixtures in the Nanogram Range and Enzyme Kinetics of Dehydrogenases Following IEF 135
19. Soft Laser Scanning Densitometer Compatible with the High Resolution Obtained by Electrofocusing 147
20. Immunocore Electrofocusing: A Separation and Detection Technique Amenable to Scanning Densitometry 153
21. Rapid, Convenient and Economical Procedures for the Determination of the Isoelectric Spectra of Proteins 155
22. A Simple Method of Choosing Optimum pH-Conditions for Electrophoresis 165
23. pH Determinations in Isoelectric Focusing with an Iridium Electrode 173
24. Anomalous Behaviour of Horseradish Peroxidase in Isoelectric Focusing 181
25. Does Lipoprotein Lipase Bind Ampholytes? 189
26. 4. Biochemical Applications
27. Zein: Macromolecular Properties, Biosynthesis and Genetic Regulation 199
28. Growth and Isoelectric Patterns of Peroxidase in Tobacco Tissue Cultures Under the Influence of Growth Regulator Systems 213
29. A Method for Simultaneous Analysis of Several Genetic Loci in Mice After Electrofocusing 221
30. Phylogenetic Differences in Isoelectric Components of Liver Acid Phosphatase and LDH Between Genera of Two Closely Related Rodent Families (Muridae and Microtidae) 233
31. Fractionation of Nucleic Acids on Isoelectric Focusing 241
32. 5. Clinical Applications
33. Polyacrylamide Gel Isoelectric Focusing of Alpha-1-Proteinase Inhibitors (α₁ Pi) Phenotypes: I. Optimal Condition, Histochemical and Proteinase Probe Studies 255
34. Polyacrylamide Gel Isoelectric Focusing (PAGIF) Applied to Alpha-1-Proteinase Inhibitor (α₁ Pi) Phenotyping: II. Immunochemical Procedures 265
35. High Resolution Phenotyping of α₁-Antitrypsins by Thin-Layer Polyacrylamide Electrofocusing 273
36. Broad-Beta Disease (Hyperlipoproteinaemia Type III): Genetics, Gene Frequency and Diagnosis without Ultracentrifugation 281
37. Screening for Abnormal Hemoglobins in the Newborn: A Highly Economic Procedure Using Isoelectric Focusing 295
38. The Crystallins of the Aging Lens from Five Species Studies by Various Methods of Thin-Layer Isoelectric Focusing 303
39. Separation of Erythrocyte Membrane Components in Electric Field. Polyacrylamide Gel Isoelectric Focusing: A Potential Method for the Diagnosis of Membrane Abnormality 313
40. Microheterogeneity of Serum Glycoproteins as Revealed by Flat-Bed Gel Isoelectric Focusing 329
41. Diagnosis of Cystic Fibrosis (CF) - Purification and Distinction Between CF Protein and Ciliary Dyskinesia Activity in CF and Asthmatic Sera Using Isoelectric Focusing and the Rabbit Tracheal Ciliary Bioassay 337
42. Microheterogeneity of Human Kininogen, Alpha- 2-HS and Alpha-1-Acid Glycoproteins by Thin- Layer Isoelectric Focusing 351
43. A Genetic Variant of Amylase from Human Parotid Saliva Detected by Isoelectric Focusing 359
44. 6. Two-Dimensional Techniques
45. Biological Applications of Two-Dimensional Gel Electrophoresis 369
46. Plant Proteins Evaluated by Two-Dimensional Methods 385
47. Two-Dimensional Separation of Lymphocyte Microsomal Membrane Proteins: Isoelectric Focusing Linked to SDS-Polyacrylamide Gel Electrophoresis 395
48. Mammalian Mitochondrial Ribosomes: Two- Dimensional Gel Electrophoresis of the 55 S- and the Corresponding Subunit Proteins 405
49. Crossed Immunoelectrofocusing for Standardization and Characterization of Fungal Antigens 413
50. 7. Preparative Separations
51. Isoelectric Focusing in Free Water 423
52. Preparative Electrofocusing in Flat-Beds of Granulated Gel - Methodological Aspects 433
53. Preparative Scale Purification of Bacterial Enzymes and Toxins by Isoelectric Focusing and Isotachophoresis 443
54. Preparative Zone Convection Electrofocusing in Helical Glass Tubes 455
55. Chromatofocusing: Isoelectric Focusing on Ion Exchangers in the Absence of an Externally Applied Potential 463
56. 8. Separation of Cells
57. New Instrumentation and Procedures for the Analysis of Mammalian Cells by Electrophoretic Techniques 469
58. Continuous-Flow Isoelectric Focusing: Further Studies on Human Red Blood Cells and on the Separation of Rat Liver Light Mitochondrial Fraction 481
59. Preparative Electrofocusing of Native and Modified Living Mammalian Cells in a Stationary Ficoll/ Sucrose Gradient 495
60. Part II. Isotachophoresis
61. 1. Analytical and Preparative Methodology
62. The Principle of Preparative Capillary Isotachophoresis 505
63. Preparative Capillary Isotachophoresis: Separation of μg Amounts of Some Human Serum Proteins 515
64. Quantitative Determination of Picomoles ADP by Means of Steady State Mixed Zones in Isotachophoresis 527
65. Thin-Layer Polyacrylamide Gel Isotachophoresis, Crossed Immuno-Isotachophoresis and Crossed Isotachophoresis-Electrofocusing 549
66. Isotachophoresis of the Cerebrospinal Fluid (CSF) Proteins: Advantage of Prior Dialysis and its Practical Realization 559
67. 2. Applications
68. Plasma Protein Fractionation in Sephadex by Isotachophoresis Using Discrete Spacers 567
69. Immuno-lsoelectric Focusing Analysis of Antibodies Fractionated by Isotachophoresis 577
70. Analytical and Preparative Isotachophoresis in Column and Flat-Bed Gels for Fractionation of Antibodies 587
71. Index 601
72. Backmatter 609