Handbook of isoelectric focusing and proteomics

Title:

Publication Information:

Amsterdam : Elsevier Academic Press, 2005

ISBN:

9780120887521

Subject Term:

Isoelectric focusing

Proteins -- Separation

Proteomics

Added Author:

Garfin, David Edward

Ahuja, Satinder, 1933-

Available:*

Library	Item Barcode	Call Number	Material Type	Item Category 1	Status
Searching... PSZ JB	30000010119412	QP519.9.I8 H36 2005	Open Access Book	Book	Searching... Unknown

Isoelectric focusing (IEF) is a high-resolution, stand-alone technique that can be used as an analytical method or tool for protein purification. The only current book on the market, the Handbook of Isoelectric Focusing and Proteomics is the ideal 'one-stop' source for germane information in this discipline. This highly practical book also contains chapters on alternative methods that may pave the way in the search for efficient techniques for fractionating and purifying proteins. Complete with the history of IEF focusing to authors' insights and practical tips, this book is a must for anyone working in proteomics.

Author Notes

Dr. Satinder Ahuja is a leading expert on improving water quality. He earned his PhD in analytical chemistry from the University of the Sciences, Philadelphia. He worked for Novartis Corp. in various leadership positions for over 25 years while simultaneously teaching and guiding research as an adjunct professor at Pace University for over 10 years. As president of Ahuja Consulting, he advises on quality issues relating to chemicals/pharmaceuticals and water. Dr. Ahuja has published numerous papers and more than 20 books. His latest books are "Chemistry and Water: The Science Behind Sustaining the World's Most Crucial Resource" (Elsevier, 2016), "Food, Energy, and Water: The Chemistry Connection" (Elsevier, 2015), and "Water Reclamation and Sustainability" (Elsevier, 2014). Others include "Comprehensive Water Quality and Purification" (Elsevier, 2013), "Monitoring Water Quality: Pollution Assessment, Analysis, and Remediation" (Elsevier, 2013), "Novel Solutions to Water Pollution" (ACS Symposium Volume, 2013), "Handbook of Water Purity and Quality" (Elsevier, 2009), and "Arsenic Contamination of Water: Mechanism, Analysis, and Remediation" (Wiley, 2008). A member of the executive committee of the Rivers of the World Foundation (ROW), Dr. Ahuja has organized numerous global symposia on improving water quality, including presentations for the American Chemical Society and UNESCO.

Pier Giorgio Righetti

Preface	p. xi
Contributors	p. xv
Incidents of Travel in IEF and IPGS	p. xvii
1 OverviewDavid E. Garfin and Satinder Ahuja
I Separations by IEF	p. 2
II Evolution and Development of IEF	p. 4
III Theory and Simulation of IEF	p. 4
IV Generation of pH Gradients	p. 5
V Slab Gel IEF	p. 5
VI Two-dimensional Gel Electrophoresis (2-DE)	p. 6
VII Practices and Pitfalls of Sample Preparation	p. 6
VIII Protein Detection and Imaging	p. 7
IX Capillary IEF	p. 7
X Preparative IEF	p. 8
XI IEF and Proteomics	p. 9
XII Chromatofocusing	p. 10
XIII Alternate Electrofocusing Methods	p. 11
XIV Summary	p. 12
References	p. 12
2 Evolution and Development of Isoelectric FocusingAkos Vegvari and Ferenc Kilar
I Introduction	p. 13
II The Rise of Electrophoresis	p. 13
III Kolin's "Isoelectric Spectra"-The Artificial pH Gradient	p. 18
IV Svensson's IEF-Vesterberg's Synthesis	p. 21
V Progress in Preparative and Analytical IEF	p. 26
VI The Immobilized pH Gradients	p. 28
VII Two-dimensional Gel Electrophoresis and Blotting of Proteins	p. 30
VIII Capillary IEF	p. 31
IX Special Features in the Practice and Theory of IEF	p. 31
X Reviews on IEF	p. 32
XI Concluding Remarks	p. 33
References	p. 34
3 Theory and Simulation of Isoelectric FocusingT.L. Sounart and P.A. Safier and J.C. Baygents
I Principles of Isoelectric Focusing	p. 41
II Numerical Simulation of IEF	p. 51
III Illustrative Simulations of IEF	p. 57
IV Summary	p. 66
References	p. 67
4 Generation of pH GradientsTom Berkelman
I Introduction	p. 69
II pH Gradients in the Early History of IEF	p. 70
III The Development of Carrier Ampholytes	p. 71
IV Practical Aspects of Carrier Ampholyte-generated pH Gradients	p. 75
V Limitations of the Carrier Ampholyte Method	p. 78
VI Early Alternative IEF Modes Not Requiring Carrier Ampholytes	p. 79
VII Immobilized pH Gradients	p. 81
VIII Use of Immobilized Buffers in Preparative IEF	p. 84
IX Practical Aspects of Immobilized pH Gradients	p. 85
References	p. 87
5 Slab Gel IEFReiner Westermeier
I Introduction	p. 93
II Equipment	p. 95
III The Gel Matrix	p. 97
IV Polyacrylamide Gels	p. 98
V Agarose Gels	p. 108
VI Dextran Gels	p. 110
VII Experimental Protocols: Polyacrylamide Slab Gel IEF	p. 111
References	p. 120
6 Two-Dimensional Gel ElectrophoresisMark P. Molloy and Michael T. McDowell
I Introduction	p. 123
II Equilibration of First Dimension IEF Gels	p. 124
III SDS-Page	p. 128
IV Protein Detection	p. 133
V Gel Reproducibility	p. 137
VI Practical Applications	p. 138
VII Advantages and Limitations of 2-DE	p. 140
VIII Summary	p. 140
References	p. 140
7 Some Practices and Pitfalls of Sample Preparation for Isoelectric Focusing in ProteomicsBen Herbert
I Introduction	p. 147
II Reduction and Alkylation	p. 150
III Beta Elimination of Cysteine	p. 152
IV Carbamylation	p. 155
V Stable Isotope Labeling-based Quantitation	p. 157
VI Sample Homogenization and Nucleic Acid Removal	p. 157
VII Membrane Proteins	p. 160
References	p. 162
8 Protein Detection and Imaging in IEF GelsWayne F. Patton
I Introduction	p. 165
II Organic Dye Staining	p. 166
III Silver Staining	p. 167
IV Reverse Staining	p. 169
V Fluorescence Staining	p. 169
VI Label-less Detection	p. 172
VII Post-translational Modification Detection	p. 172
VIII Acquiring Images from Stained Gels	p. 173
IX Conclusion	p. 176
Acknowledgments	p. 176
References	p. 176
9 Capillary Isoelectric FocusingTim Wehr
I Introduction	p. 181
II Sample Preparation	p. 183
III Ampholyte Selection and Sample Introduction	p. 185
IV Focusing	p. 186
V Mobilization Techniques	p. 187
VI Capillary Selection	p. 193
VII Minimizing Protein Precipitation	p. 195
VIII Internal Standards for cIEF	p. 195
IX Imaging cIEF	p. 196
X cIEF-Mass Spectrometry	p. 197
XI cIEF In Microchannels	p. 199
XII Applications of cIEF	p. 200
References	p. 205
10 Free-Flow Isoelectric FocusingPeter J.A. Weber and Gerhard Weber and Christoph Eckerskorn and Ulrich Schneider and Anton Posch
I Introduction	p. 211
II Principle of EFE	p. 212
III Instrumentation	p. 220
IV Applications	p. 231
V Summary	p. 239
References	p. 239
11 Isoelectric Focusing and ProteomicsMelanie Y. White and Stuart J. Cordwell
I Introduction	p. 247
II The Proteomics Workflow	p. 249
III IEF for Prefractionation	p. 252
IV IEF in Two-dimensional Electrophoresis	p. 254
V Conclusions	p. 259
Glossary	p. 259
Acknowledgments	p. 259
References	p. 260
12 ChromatofocusingDavid Anderson
I Introduction	p. 265
II Conventional Chromatofocusing (Internal pH Gradient Generation)	p. 267
III Gradient Chromatofocusing (External pH Gradient Generation)	p. 277
IV Performance Characteristics	p. 283
V Applications	p. 288
References	p. 290
13 Alternative Electrofocusing MethodsCornelius F. Ivory
I Introduction	p. 297
II Theory	p. 301
III Results	p. 311
IV Discussion	p. 315
V Conclusion	p. 317
Acknowledgments	p. 317
References	p. 318
Index	p. 321

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