- Email: [email protected]
Get electrophoresis of proteins (edited by H.J. Dunn)
38
measured and used to evaluate the performance of a given detector. Emphasis is put on the fact that the detector is only part of the chromatographic system and that other elements also have to be optimized in order to obtain optimum signal to noise performance. The description of the various types of detectors and the underlying physical and chemical phenomena is extensive and gives an excellent overview of both commonly used detectors and those which are of more academic interest. In spite of their practical significance the diode array detector and the fluorometric detector are only treated briefly. The nu-
trends in analytical chemistry, vol. 7, no. I, 1988 .
merous possibilities these detectors render for the improvement of selectivity and sensitivity are not dealt with in much detail. A real wealth of information for the practitioner is the chapter on the selection of the appropriate detector with practical hints on detector operation, particularities of quantitative and qualitative analysis and computer data processing. Each chapter ends with a comprehensive synopsis both summarizing the essential points and highlighting the practically relevant elements. The literature citations appear to be complete. The book can be recommended to both the novice and the experienced
After the flood Gel Electrophoresis of Proteins, edited by M. .I. Dunn, Wright, 1986, f 45.00 (407 pages) ZSBN 0-72360882-2
The recent deluge of books on the subject of electrophoresis has made it impossible for every investigator who uses this technique to consider purchasing each one. The quality of these offerings spans a considerable spectrum, thus a decision to buy must be carefully made. In this reviewer’s opinion, the current volume is worthy of serious consideration. The first chapter is concerned with theory (Schafer-Nielsen). This is a fresh approach, discussing various electrophoretic processes from the point of view of the steady states which are produced. The fundamental physical principles which contribute to the theory are presented in clear manner. The chapter lacks two things: a discussion of the role of diffusion in the maintenance of electrophoretic steady states and a presentation of the theory of zone electrophoresis, which is perhaps the most widely used variant, but is omitted because it lacks any characteristic steady state behavior.
The second chapter is a very thorough discussion of single dimensional separations (Rothe and Maurer) utilizing zone electrophoretic principles. There is also a section on affinity electrophoresis. Separations under denaturing and non-denaturing conditions, in uniform, discontinuous and gradient gels using continuous and discontinuous buffer systems are all covered in detail. This is an excellent chapter, augmented with numerous tables to organize the large amount of information presented and with an extensive reference list. The only shortcoming is the absence of a discussion of detergents other than SDS. The next chapter is concerned with conventional isoelectric focusing (IEF) and focusing in immobilized pH gradients (IPGs) (Righetti, Gelfi and Gianazza). The presentation of conventional IEF is a good one and is complimented with a section on the generation of pH mobility curves for proteins and a discussion of their various uses. Both the IPG and IEF sections contain much methodological information. Perhaps the most valuable aspects of this chapter are a good discussion of artifacts and a troubleshooting guide.
chromatographer. The former will get an excellent introduction to all aspects of detection in liquid chromatography and will find a lot of practical hints and advice on the operation of the various detector types; the latter will be amazed to see the vast amount of information which has been compiled around the subject of detection in modern liquid chromatography during the past 20 years. K. P. HUPE
K. P. Hupe is at Hewlett-Packard GmbH, Analytische Messtechnic, Postfach 1280, D-751 7 Waldbronn 2, F. R. G.
Chapter 4, on two dimensional separations (Dunn and Burghes), begins with an historical review of the area followed by a description of methods employing no denaturants for the isolation of biologically active proteins. Also included is a discussion of 2-D methods using zone electrophoresis in both dimensions. The description of techniques is quite extensive and this excellent chapter also contains a tabular summary of the optimal features of a 2-D system with comparisons drawn to the popular O’Farrell method. The presentation of immunoelectrophoretic methods (Heegaard and Bog-Hansen) covers all such techniques in. current use. The emphasis is not as much on detailed presentation of methods and procedures but rather on description of the basic precepts, applicability to particular problems and interpretation of results. There is an extensive discussion of the use of modified antigens and also of the characterization of antigens. The chapter concerning peptide mapping (Gooderham) is very short and is little more than a thorough introduction to the concept. Protein staining using organic dyes and silver is very nicely covered by Merril, Harasewych and Harrington. A detailed discussion of the detection of radiolabelled proteins, enzyme
39
trends iqwalytical chemktry, vol. 7, no. 1,1988
stains, the detection of proteinbound trace elements and detection after electroblotting are also included. Missing from this otherwise excellent chapter is some mention of other metallic stains. The final chapter addresses the looming problem of capturing, storing, retrieving and comparing the very large amounts of data present in 2-D gels (Spragg, Amess, Jones and Ramasamy). It is oriented to the investigator who may
wish to develop a complete system, outlining the many options and obstacles inherent in such a task. Unfortunately, there is little information for those interested in comparison of currently available technology. This volume contains no chapter dedicated to agarose gels, rather this information is found throughout the book, which is not a serious limitation. In summary, this compilation
contains several excellent chapters, falls in the high quality end of the aforementioned spectrum and would make a valuable addition to any electrophoretic library. R. A. MOSHER Richard
A. Mosher
is Associate Director
ofthe Center for Separation Science, University of Arizona, Bldg. 20; Room 156, Tuczon, AZ 85721, U.S.A.
THE PROTEIN SEQUENCING APPRENTICE A program which simulates the sequencing of a protein
By A.R. Place and T.G. Schmidt, IRL Press, 1986. Technical and price specifications:
Computer Operating System Language Minimum Memory Storage Medium Required peripherals Order Ref. No. Price (including manuals)
IBM PC/XT/AT PC/MS-DOS 2.0+ Turbo Pascal 256K RAM 5% in. diskette 3563 diskette drive and colour graphics adaptor ISBN 0 947946 57 8 US$99.00, f 65.00 (plus VAT in U.K.)
‘The Protein Sequencing Apprentice’ is a simulation that enables you to apply over 20 techniques to your peptide sample. This sample may have a known sequence that you have entered or an unknown sequence, generated by the computer for a peptide length that you have specified. You may opt for the ‘practice’ mode when you are given 10 000 nmol of sample or for the ‘test’ mode with only 100 nmol. I would have liked something in between because I often ran out of sample in the ‘test’ mode. However, the ‘show sequence’ command at least allows one to check partial achievement . It is possible to interrupt a session, store the data and resume later. You can invoke ‘inventory’ to show the cleavages you have performed and how much of the starting materi-
al remains. The ‘Notes for Students’ describes in clear detail the assumptions made in programming the various techniques, for example the residues destroyed during acid or base hydrolysis. The same information for any technique, can be shown on the screen when requested by the command ‘mechanism’. The techniques available are: acid hydrolysis, base hydrolysis, isoelectric point, performic acid oxidation, end group analysis (N terminal residue), carboxypeptidases (either the residues liberated after a single time or a time course, which, of course, takes more peptide), Edman degradation (either ‘automatic’, when you can specify how much sample and the results are shown as HPLC traces or ‘manual’ with TLC display), pH 2.5 hydrolysis, hydroxylamine, cyano-
gen bromide, staphylococcal protease, cyanolysis, trypsin, clostripain, iodosobenzoate, N-bromosuccinimide, pepsin, thermolysin and partial acid hydrolysis (which allows one to follow in Sanger’s footsteps, but the program does not permit cheating by using Edman on the fragments). Clearly this is an extensive range of techniques which allow a considerable degree of realism in selecting a strategy for analysis. The publisher’s claim that ‘Students can use this software to recreate the effects of laboratory techniques . . . ’ is quite correct but for an undergraduate student to explore all of them would take many hours and he would probably learn more than he needs to know about sequencing. It could be useful for a postgraduate student serving a protein sequencing
measured and used to evaluate the performance of a given detector. Emphasis is put on the fact that the detector is only part of the chromatographic system and that other elements also have to be optimized in order to obtain optimum signal to noise performance. The description of the various types of detectors and the underlying physical and chemical phenomena is extensive and gives an excellent overview of both commonly used detectors and those which are of more academic interest. In spite of their practical significance the diode array detector and the fluorometric detector are only treated briefly. The nu-
trends in analytical chemistry, vol. 7, no. I, 1988 .
merous possibilities these detectors render for the improvement of selectivity and sensitivity are not dealt with in much detail. A real wealth of information for the practitioner is the chapter on the selection of the appropriate detector with practical hints on detector operation, particularities of quantitative and qualitative analysis and computer data processing. Each chapter ends with a comprehensive synopsis both summarizing the essential points and highlighting the practically relevant elements. The literature citations appear to be complete. The book can be recommended to both the novice and the experienced
After the flood Gel Electrophoresis of Proteins, edited by M. .I. Dunn, Wright, 1986, f 45.00 (407 pages) ZSBN 0-72360882-2
The recent deluge of books on the subject of electrophoresis has made it impossible for every investigator who uses this technique to consider purchasing each one. The quality of these offerings spans a considerable spectrum, thus a decision to buy must be carefully made. In this reviewer’s opinion, the current volume is worthy of serious consideration. The first chapter is concerned with theory (Schafer-Nielsen). This is a fresh approach, discussing various electrophoretic processes from the point of view of the steady states which are produced. The fundamental physical principles which contribute to the theory are presented in clear manner. The chapter lacks two things: a discussion of the role of diffusion in the maintenance of electrophoretic steady states and a presentation of the theory of zone electrophoresis, which is perhaps the most widely used variant, but is omitted because it lacks any characteristic steady state behavior.
The second chapter is a very thorough discussion of single dimensional separations (Rothe and Maurer) utilizing zone electrophoretic principles. There is also a section on affinity electrophoresis. Separations under denaturing and non-denaturing conditions, in uniform, discontinuous and gradient gels using continuous and discontinuous buffer systems are all covered in detail. This is an excellent chapter, augmented with numerous tables to organize the large amount of information presented and with an extensive reference list. The only shortcoming is the absence of a discussion of detergents other than SDS. The next chapter is concerned with conventional isoelectric focusing (IEF) and focusing in immobilized pH gradients (IPGs) (Righetti, Gelfi and Gianazza). The presentation of conventional IEF is a good one and is complimented with a section on the generation of pH mobility curves for proteins and a discussion of their various uses. Both the IPG and IEF sections contain much methodological information. Perhaps the most valuable aspects of this chapter are a good discussion of artifacts and a troubleshooting guide.
chromatographer. The former will get an excellent introduction to all aspects of detection in liquid chromatography and will find a lot of practical hints and advice on the operation of the various detector types; the latter will be amazed to see the vast amount of information which has been compiled around the subject of detection in modern liquid chromatography during the past 20 years. K. P. HUPE
K. P. Hupe is at Hewlett-Packard GmbH, Analytische Messtechnic, Postfach 1280, D-751 7 Waldbronn 2, F. R. G.
Chapter 4, on two dimensional separations (Dunn and Burghes), begins with an historical review of the area followed by a description of methods employing no denaturants for the isolation of biologically active proteins. Also included is a discussion of 2-D methods using zone electrophoresis in both dimensions. The description of techniques is quite extensive and this excellent chapter also contains a tabular summary of the optimal features of a 2-D system with comparisons drawn to the popular O’Farrell method. The presentation of immunoelectrophoretic methods (Heegaard and Bog-Hansen) covers all such techniques in. current use. The emphasis is not as much on detailed presentation of methods and procedures but rather on description of the basic precepts, applicability to particular problems and interpretation of results. There is an extensive discussion of the use of modified antigens and also of the characterization of antigens. The chapter concerning peptide mapping (Gooderham) is very short and is little more than a thorough introduction to the concept. Protein staining using organic dyes and silver is very nicely covered by Merril, Harasewych and Harrington. A detailed discussion of the detection of radiolabelled proteins, enzyme
39
trends iqwalytical chemktry, vol. 7, no. 1,1988
stains, the detection of proteinbound trace elements and detection after electroblotting are also included. Missing from this otherwise excellent chapter is some mention of other metallic stains. The final chapter addresses the looming problem of capturing, storing, retrieving and comparing the very large amounts of data present in 2-D gels (Spragg, Amess, Jones and Ramasamy). It is oriented to the investigator who may
wish to develop a complete system, outlining the many options and obstacles inherent in such a task. Unfortunately, there is little information for those interested in comparison of currently available technology. This volume contains no chapter dedicated to agarose gels, rather this information is found throughout the book, which is not a serious limitation. In summary, this compilation
contains several excellent chapters, falls in the high quality end of the aforementioned spectrum and would make a valuable addition to any electrophoretic library. R. A. MOSHER Richard
A. Mosher
is Associate Director
ofthe Center for Separation Science, University of Arizona, Bldg. 20; Room 156, Tuczon, AZ 85721, U.S.A.
THE PROTEIN SEQUENCING APPRENTICE A program which simulates the sequencing of a protein
By A.R. Place and T.G. Schmidt, IRL Press, 1986. Technical and price specifications:
Computer Operating System Language Minimum Memory Storage Medium Required peripherals Order Ref. No. Price (including manuals)
IBM PC/XT/AT PC/MS-DOS 2.0+ Turbo Pascal 256K RAM 5% in. diskette 3563 diskette drive and colour graphics adaptor ISBN 0 947946 57 8 US$99.00, f 65.00 (plus VAT in U.K.)
‘The Protein Sequencing Apprentice’ is a simulation that enables you to apply over 20 techniques to your peptide sample. This sample may have a known sequence that you have entered or an unknown sequence, generated by the computer for a peptide length that you have specified. You may opt for the ‘practice’ mode when you are given 10 000 nmol of sample or for the ‘test’ mode with only 100 nmol. I would have liked something in between because I often ran out of sample in the ‘test’ mode. However, the ‘show sequence’ command at least allows one to check partial achievement . It is possible to interrupt a session, store the data and resume later. You can invoke ‘inventory’ to show the cleavages you have performed and how much of the starting materi-
al remains. The ‘Notes for Students’ describes in clear detail the assumptions made in programming the various techniques, for example the residues destroyed during acid or base hydrolysis. The same information for any technique, can be shown on the screen when requested by the command ‘mechanism’. The techniques available are: acid hydrolysis, base hydrolysis, isoelectric point, performic acid oxidation, end group analysis (N terminal residue), carboxypeptidases (either the residues liberated after a single time or a time course, which, of course, takes more peptide), Edman degradation (either ‘automatic’, when you can specify how much sample and the results are shown as HPLC traces or ‘manual’ with TLC display), pH 2.5 hydrolysis, hydroxylamine, cyano-
gen bromide, staphylococcal protease, cyanolysis, trypsin, clostripain, iodosobenzoate, N-bromosuccinimide, pepsin, thermolysin and partial acid hydrolysis (which allows one to follow in Sanger’s footsteps, but the program does not permit cheating by using Edman on the fragments). Clearly this is an extensive range of techniques which allow a considerable degree of realism in selecting a strategy for analysis. The publisher’s claim that ‘Students can use this software to recreate the effects of laboratory techniques . . . ’ is quite correct but for an undergraduate student to explore all of them would take many hours and he would probably learn more than he needs to know about sequencing. It could be useful for a postgraduate student serving a protein sequencing