Laboratory techniques in biochemistry and molecular biology / Vol. 22: Techniques in free radical research / Catherine A. Rice-Evans, Anthony T. Diplock, Martyn C.R. Symons

Free radical species are generally short-lived due to their high reactivity and thus direct measurement and identification are often impossible. ESR is the only technique which has the potential for direct detection of radicals but in biological systems even these must be trapped by a spin-trapping agent. Thus most investigations involve recognition of indicators of the presence of radicals in vivo or "FOOTPRINTS" of radical-mediated damage. Techniques in Free Radical Research assembles and critically assesses the most relevant and reliable experimental approaches used towards the measurement of radicals and radical-mediated damage in chemical systems, in cells and in tissues under the following six headings: a) Footprints of DNA damage, b) Footprints of protein damage, c) Footprints of lipid peroxidation, d) Footprints of antioxidant consumption, e) Footprints via indirect radical assays, and f) Footprints via the availability of transition metal complexes. v. 1. pt. 1. Electrophoresis of proteins in polyacrylamide and starch gels, by A.H. Gordon.- pt. 2. An introduction to gel chromatography, by L. Fischer.- pt. 3. Immunochemical techniques for the identification and estimation of macromolecules, by J. Clausen. v. 2. pt. 1. Automated enzyme assays, by D.B. Roodyn.- pt. 2. Cellulosic ion exchangers, by E.A. Peterson. v. 3. pt. 1. Determination of sequences in RNA, by G.G. Brownlee.- pt. 2. Techniques of lipidology, by M. Kates. v. 4. pt. 1. Chemical modification of proteins, by A.N. Glazer, R.J. DeLange and D.S. Sigman.- pt. 2. Separation methods for nucleic acids and oligonucleotides, by H. Gould and H.R. Matthews. v. 5. pt. 1. Techniques of sample preparation of liquid scintillation counting, by B.W. Fox.- Isoelectric focusing, by P.G. Righetti and J.W. Drysdale. v. 6. pt. 1. Density gradient centrifugation, by R. Hinton and M. Dobrota.- pt. 2. An introduction to radioimmunoassay and related techniques, by T. Chard. v. 7, pt. 1. Preparation and characterisation of mammalian plasma membranes, by W.H. Evans. pt. 2. An introduction to affinity chromatography, by C.R. Lowe. v. 8. Cell culture for biochemists, by R.L.P. Adams. v. 9. Sequencing of proteins and peptides, by G. Allen. v. 10. DNA sequencing, [by] J. Hindley. v. 11. Isoeletric focusing: theory, methodology and applications, [by] P. G. Righetti. v. 12. Photogenerated reagents in biochemistry and molecular biology, [by] H. Bayley. v. 13. Monoclonal antibody technology: the production and characterization of rodent and human hybridomas, [by] A.M. Campbell. v. 14. A guidebook to lipoprotein technique, [by] G.L. Mills, P.A. Lane, and P.K. Weech. v. 15. Practice and theory of enzyme immunoassays, [by] P. Tijssen. v. 16. Glycoprotein and proteoglycan techniques, [by] J.G. Beeley. v. 17. Applications of hplc in biochemistry, [by] A. Fallon, R.F.G. Booth and L.D. Bell. v. 18. Methods of cell separation, [by] P.T. Sharpe. v. 19. Synthetic polypeptides as antigens [by] M.H.V. Van Regenmortel [et al.]. v. 20. Immobilized pH gradients: theory and methodology, by] P. G. Righetti. v. 21. Isolated hepatocytes preparation, properties and applications by M.N. Berry, A.M. Edwards [and] G.J. Barritt. v. 22. Techniques in free radical research, edited by R.H. Burdon [and] P.H. van Knippenberg. v. 23. Monoclonal antibody and immunosensor technology, edited by P.C. van der Vliet. v. 24. Hybridization with nucleic acid probes.- pt. 1. Theory and nucleic acid preparation.- pt. 2. Probe labeling and hybridization techniques. v. 25. Dry chemistry. v. 26. Analysis of RNA-protein complexes in Vitro v. 27. Ultrasensitive and rapid enzyme immunoassay / Eiji Ishikawa v. 28. Synthetic peptides as antigens / M.H.V. Van Regenmortel, S. Muller. v. 29. Cancer metastasis: in vitro and in vivo experimental approaches v. 30. Inositol phospholipid metabolism and phosphatidyl inositol kinases v. 32. Magnetic cell separation. 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: 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.

Free radical species are generally short-lived due to their high reactivity and thus direct measurement and identification are often impossible. ESR is the only technique which has the potential for direct detection of radicals but in biological systems even these must be trapped by a spin-trapping agent. Thus most investigations involve recognition of indicators of the presence of radicals in vivo or "FOOTPRINTS" of radical-mediated damage.

Techniques in Free Radical Research assembles and critically assesses the most relevant and reliable experimental approaches used towards the measurement of radicals and radical-mediated damage in chemical systems, in cells and in tissues under the following six headings: a) Footprints of DNA damage, b) Footprints of protein damage, c) Footprints of lipid peroxidation, d) Footprints of antioxidant consumption, e) Footprints via indirect radical assays, and f) Footprints via the availability of transition metal complexes.
Content: Edited By Page iii Copyright page Page iv Preface Pages v-vii Catherine Rice-Evans, Anthony T. Diplock, Martyn C.R. Symons Chapter 1 Introduction to free radicals Pages 1-18 Chapter 2 Mechanisms of radical production Pages 19-50 Chapter 3 The detection and characterization of free radical species Pages 51-100 Chapter 4 Transition metal complexes as sources of radicals Pages 101-124 Chapter 5 Investigation of the consequences of free radical attack on lipids Pages 125-184 Chapter 6 Assay of antioxidant nutrients and antioxidant enzymes Pages 185-206 Chapter 7 Detection of protein structural modifications induced by free radicals Pages 207-235 Chapter 8 Determination of radical damage to DNA Pages 237-252 Appendix A Pages 253-255 Appendix B - Methods for isolation of low density lipoproteins and erythrocyte membranes and cytoskeletons from blood Pages 257-264 References Pages 265-278 Subject index Pages 279-291 Free radical species are generally short-lived due to their high reactivity and thus direct measurement and identification are often impossible. ESR is the only technique which has the potential for direct detection of radicals but in biological systems even these must be trapped by a spin-trapping agent. Thus most investigations involve recognition of indicators of the presence of radicals in vivo or andquot;FOOTPRINTSandquot; of radical-mediated damage. Techniques in Free Radical Research assembles and critically assesses the most relevant and reliable experimental approaches used towards th