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Gel electrophoresis of proteins
251
Journal of Chonautography,414 (1987) 251-262
Biomedical Applications Elsevier Science Publishers B.V., Am&dam
-
printed in The Netherlands
CHROMBIO. 3410
Book Review Gel eZ&rophoresis ofproteins, editedby M.J. Dunn, Wright,Bristol,1986,xiii+ 407
pp, price .&COO,ISBN o-7236-0882-2.
Apart from the rapid expansionof electromigrationseparationsof nucleicacids and their fragments,proteins remain the dominant materialsto be separatedby electrophoresis.The developmentof this field is relativelyfast and, perhaps,three aspectscan currentlybe highlightedwith regardto proteins: the side-by-sideseparationof individualmembersof very complex mixtures,the isolation of a single protein from a very complex mixture and quantitation of the results. How does the present volume cover these topics? A general introduction to electromigrationtechniques that offers a well balanced amount of theory and that explains lucidly the basic differences between individual techniques (zone electrophoresis, isotachophoresis and isoelectric focusing) is hidden behind a rather cumbersome title, Steady-state gel electrophoresis systems (Shafer-Nielsen). The next chapter, by Rothe and Maurer, deals with one-dimensionalseparationsof both native and denaturedproteins in polyacrylamidegel.This chaptercontains a numberof particularlyvaluabletables regarding,e.g., the application of individualproteins as molecular weight standards buffer systems used for protein separations, and is generallypleasant to read. It also contains a section on affinity electrophoresis.The reviewer’sobjections to this chapter are fairly minor and concern both the subject and the formal presentation.It would have certainlybeen worth discussingalso detergentsother than SDS here. Why is there such emphasison membraneproteins that they are given a special section (p. 112)? Nobody would deny that membrane proteins representan importantcategory,but other categoriesof proteinsare surelyequally important.The readerwouldprobably requiremore detailedinformationon other protein types or, if the book had been kept on a more generallevel, he would not havemindedthe omissionof thesefew pages,whichsimplydo not fit here.Another aspect that is disliked is the omission of headingsfrom Tables 2.9 and 2.10 (pp. 80and81). The third chapter, by Righetti, Gelfi and Gianaxza, describes conventional isoelectricfocusingand immobilizedpH gradients.In the introductionthe authors state that because of the recent explosion of the number of books dealing with electrophoretictechniques,they “will have to strugglehard to give a modern and freshview of the two techniques.” Their chaptercan be consideredto be a success;
252
it gives an overall view of the topic, with enough technical details to be useful to a newcomer, together with a table of “disasters” that may serve aa a guide for even experienced workers to improve their results. The chapter by Dunn and Burghes on high-resolution twodimensional polyacrylamide gel electrophoresis is superb. Nevertheless, one criticism, similar to that mentioned afore for the separation of membrane proteins, can be made, namely that title of the last section, BD-PAGE and the study of human genetic disorders, seemingly does not fit in the overall scope of the chapter. On reading it, one’s doubts disappear, but it would have been better to have had a more accurate title. Immunoelectrophoretic methods are reviewed by Heegaard and Bog-Hansen. This chapter provides a well balanced proportion of theory, practical hints and applications. All the main procedures are surveyed and the chapter is detailed enough to serve as a laboratory manual (in fact this also holds for most of the other chapters). Peptide mapping is certainly a powerful tool for elucidating protein structures. Blectromigration methods relevant to this are reviewed by K. Gooderham; the word that would describe best this part of the book is superficial. The chapter covers a mere ten pages, of which one page is devoted to enzymic cleavage procedures and half a page to chemical cleavage of protein molecules. These topics clearly require more space and a deeper insight. Incidentally, when the author states that, e.g. Staphylococcus aureus protease cleaves proteins at Asp and Glu, it is not obvious whether he means the C- or the N-terminus of the amino acid (Table 6.1, p. 317) ; for a newcomer such lack of clarity can be confusing. The last two’chapters are by Merril, Harasewych and Harrington on Protein staining and detection methods and by Spragg, Imes, Jones and Bamasamy on Quantifying patterns from two-dimensional PAGE. Here current detection procedures such as silver staining, enzyme staining, detection of radioactive proteins and detection of protein-bound trace elements are reviewed, together with techniques for quantitative evaluation, i.e., scanning procedures, data collection, programming languages, computerization and numerical analysis. The whole volume is completed with a well prepared subject index. To answer the question in the first paragraph, the book covers all the major topics more than adequately. The single weak point is peptide mapping, which can be considered to be on the borderline of the problems with which the book is concerned. The few objections raised here represent minor errors rather than serious drawbacks. The book is, in reviewer’s opinion, worthy of a place on the protein chemist’s bookshelves and, appears to be a recommendable purchase. Prague (Czechoslovakia)
2. DEYL
Journal of Chonautography,414 (1987) 251-262
Biomedical Applications Elsevier Science Publishers B.V., Am&dam
-
printed in The Netherlands
CHROMBIO. 3410
Book Review Gel eZ&rophoresis ofproteins, editedby M.J. Dunn, Wright,Bristol,1986,xiii+ 407
pp, price .&COO,ISBN o-7236-0882-2.
Apart from the rapid expansionof electromigrationseparationsof nucleicacids and their fragments,proteins remain the dominant materialsto be separatedby electrophoresis.The developmentof this field is relativelyfast and, perhaps,three aspectscan currentlybe highlightedwith regardto proteins: the side-by-sideseparationof individualmembersof very complex mixtures,the isolation of a single protein from a very complex mixture and quantitation of the results. How does the present volume cover these topics? A general introduction to electromigrationtechniques that offers a well balanced amount of theory and that explains lucidly the basic differences between individual techniques (zone electrophoresis, isotachophoresis and isoelectric focusing) is hidden behind a rather cumbersome title, Steady-state gel electrophoresis systems (Shafer-Nielsen). The next chapter, by Rothe and Maurer, deals with one-dimensionalseparationsof both native and denaturedproteins in polyacrylamidegel.This chaptercontains a numberof particularlyvaluabletables regarding,e.g., the application of individualproteins as molecular weight standards buffer systems used for protein separations, and is generallypleasant to read. It also contains a section on affinity electrophoresis.The reviewer’sobjections to this chapter are fairly minor and concern both the subject and the formal presentation.It would have certainlybeen worth discussingalso detergentsother than SDS here. Why is there such emphasison membraneproteins that they are given a special section (p. 112)? Nobody would deny that membrane proteins representan importantcategory,but other categoriesof proteinsare surelyequally important.The readerwouldprobably requiremore detailedinformationon other protein types or, if the book had been kept on a more generallevel, he would not havemindedthe omissionof thesefew pages,whichsimplydo not fit here.Another aspect that is disliked is the omission of headingsfrom Tables 2.9 and 2.10 (pp. 80and81). The third chapter, by Righetti, Gelfi and Gianaxza, describes conventional isoelectricfocusingand immobilizedpH gradients.In the introductionthe authors state that because of the recent explosion of the number of books dealing with electrophoretictechniques,they “will have to strugglehard to give a modern and freshview of the two techniques.” Their chaptercan be consideredto be a success;
252
it gives an overall view of the topic, with enough technical details to be useful to a newcomer, together with a table of “disasters” that may serve aa a guide for even experienced workers to improve their results. The chapter by Dunn and Burghes on high-resolution twodimensional polyacrylamide gel electrophoresis is superb. Nevertheless, one criticism, similar to that mentioned afore for the separation of membrane proteins, can be made, namely that title of the last section, BD-PAGE and the study of human genetic disorders, seemingly does not fit in the overall scope of the chapter. On reading it, one’s doubts disappear, but it would have been better to have had a more accurate title. Immunoelectrophoretic methods are reviewed by Heegaard and Bog-Hansen. This chapter provides a well balanced proportion of theory, practical hints and applications. All the main procedures are surveyed and the chapter is detailed enough to serve as a laboratory manual (in fact this also holds for most of the other chapters). Peptide mapping is certainly a powerful tool for elucidating protein structures. Blectromigration methods relevant to this are reviewed by K. Gooderham; the word that would describe best this part of the book is superficial. The chapter covers a mere ten pages, of which one page is devoted to enzymic cleavage procedures and half a page to chemical cleavage of protein molecules. These topics clearly require more space and a deeper insight. Incidentally, when the author states that, e.g. Staphylococcus aureus protease cleaves proteins at Asp and Glu, it is not obvious whether he means the C- or the N-terminus of the amino acid (Table 6.1, p. 317) ; for a newcomer such lack of clarity can be confusing. The last two’chapters are by Merril, Harasewych and Harrington on Protein staining and detection methods and by Spragg, Imes, Jones and Bamasamy on Quantifying patterns from two-dimensional PAGE. Here current detection procedures such as silver staining, enzyme staining, detection of radioactive proteins and detection of protein-bound trace elements are reviewed, together with techniques for quantitative evaluation, i.e., scanning procedures, data collection, programming languages, computerization and numerical analysis. The whole volume is completed with a well prepared subject index. To answer the question in the first paragraph, the book covers all the major topics more than adequately. The single weak point is peptide mapping, which can be considered to be on the borderline of the problems with which the book is concerned. The few objections raised here represent minor errors rather than serious drawbacks. The book is, in reviewer’s opinion, worthy of a place on the protein chemist’s bookshelves and, appears to be a recommendable purchase. Prague (Czechoslovakia)
2. DEYL