Immobilized pH Gradients: Theory and MethodologyImmobilized 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: detailed treatment of all the chemistry of the Immobiline chemicals; theory of pH gradient generation (computer simulations, tables with all the possible pH recipes); all analytical aspects of IPGs, including staining techniques, blotting etc.; two-dimensional maps, with a detailed treatise of advantages and limitations; preparative aspects of IPGs, including comparison with other preparative electrophoretic techniques; some examples of applications, including genetic and forensic analysis, blood polymorphism etc. The book is extensive and up-to-date, while also extensively covering the theory. Clearly written, with easily accessible information, the volume is a worthy asset to the Lab. Techniques series. |
Contents
| 1 | |
| 5 | |
Chapter 2 Theory | 53 |
Chapter 3 Analytical IPGs | 117 |
Chapter 4 Twodimensional 2D maps | 181 |
Chapter 5 Preparative aspects of immobilized pH gradients | 257 |
Chapter 6 Some applications of IPGs | 317 |
Maestro Volta | 373 |
| 375 | |
| 395 | |
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Common terms and phrases
2-D maps acrylamide acrylic acid addition agarose alkaline Immobilines amino acid Ampholine ampholytes amphoteric analysis anodic antibody apparatus applied bands blotting Buffer concentrations buffering Immobiline buffering power CA-IEF carrier ampholytes cassette cathodic cell Cham 2 Cham containing Coomassie detection detergent dimension electrode electrophoretic elution enzyme equilibration flow chamber focusing Gel Bond PAG Gelfi Gianazza Görg gradient mixer hydrophobic Immobiline Immobiline chemicals Immobiline gel Immobiline pK immobilized pH gradients ionic strength IPG gel IPG gel strip IPG interval IPG matrix IPG pH IPG run IPG technique isoelectric isoelectric point membrane method mixture molarity molecules ovalbumin oxidation permission from Elsevier permission of Elsevier pH gradient pH interval pH range pH values plate polymerization procedure protein load protein zone Righetti salt sample application separation shown in Fig silver staining solubility spots Table tion titrant tubes urea variants voltage washing