Immobilized pH Gradients : Theory and Methodology
Pier Giorgio. Righetti
Bok Engelsk 1990 · Electronic books.
| Annen tittel | |
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| Utgitt | Burlington : : Elsevier Science, , 1990.
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| Omfang | 1 online resource (415 p.)
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| Opplysninger | Description based upon print version of record.. - Front Cover; IMMOBILIZED pH GRADIENTS: THEORY AND METHODOLOGY; Copyright Page; Contents; List of abbreviations; Acknowledgments; Introduction; Chapter 1. The chemicals; 1.1 Isoelectric focusing: an evolving concept; 1.2 Comparison between carrier ampholyte and Immobiline chemicals; 1.3 Structure and synthesis of acidic Immobiline; 1.4 Structure and synthesis of basic Immobiline; 1.5 Physico-chemical properties of Immobiline; 1.6 Problems with the Immobiline chemicals; 1.7 Storage of Immobiline: a newgeneration; 1.8 Conclusions; Chapter 2. Theory; 2.1 Per aspera ad astra. - 2.2 Narrow and ultra-narrow pH gradients2.3 Extended pH gradients; 2.4 How to deal with experimental errors; 2.5 Ionic strength (I); 2.6 Buffering capacity (ß); 2.7 Conductivity; 2.8 Electroendosmosis; 2.9 The limits of IPGs: how to break barriers; 2.10 Resolving power. Rilbe's equation; 2.11 Alternatives for buffer focusing; 2.12 Conclusions; Chapter 3. Analytical IPGs; 3.1 General considerations; 3.2 Equipment; 3.3 The polyacrylamide gel matrix; 3.4 Gel preparation and electrophoresis; 3.5 General experimental considerations; 3.6 Blotting from Immobiline gels; 3.7 Mixed-bed, CA-IPG gels. - 3.8 Effect of salts on protein patterns in IPG gels3.9 General staining procedures; 3.10 Trouble shooting; 3.11 Conclusions; Chapter 4. Two-dimensional (2-D) maps; 4.1 Introduction; 4.2 Sample preparation; 4.3 Sample solubilization; 4.4 Choice of detergent; 4.5 Radiolabelling techniques; 4.6 The first (IPG) dimension; 4.7 The second dimension; 4.8 Reproducibility of spot positions in 2-D maps; 4.9 Computer data acquisition; 4.10 Silver staining; 4.11 Blotting of spots from 2-D maps; 4.12 General experimental aspects; 4.13 Trouble shooting; 4.14 Examples of some applications. - 4.15 To be or not to be?Chapter 5. Preparative aspects of immobilized pH gradients; 5.1 Introduction; 5.2 Theory; 5.3 Methodology; 5.4 Immobiline canal technique; 5.5 Reversible polyacrylamide gels; 5.6 Segmented immobilized pH gradients; 5.7 Alternative preparative methods; 5.8 Conclusions; Chapter 6. Some applications of IPGs; 6.1 Analysis of hemoglobins; 6.2 Polymorphism of human proteins; 6.3 Zymograms; 6.4 Focusing at pH extremes; 6.5 Conclusions. On protein heterogeneity; Chapter 7. Post scriptum: Maestro Volta; References; Subject Index. - Immobilized pH gradients (IPG) represent the most advanced development of isoelectric focusing (IEF). Originally developed to overcome all the problems of IEF in soluble amphoteric buffers (CA) (such as pH gradient instability, complexation with CA chemicals, unreproducibility of pH gradients, protein precipitation at the pI), it turned out to be an entirely new technique, quite different in principle and operation from conventional IEF. The book is thus meant to bring the reader up to date with this fast developing field. The book is divided into six chapters containing information on
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| Dewey | |
| ISBN | 0444813012. - 0444813152
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