By Rosaria P. Haugland, Mahesh K. Bhalgat (auth.), Robert J. McMahon (eds.)
For researchers at the present time, the avidin-biotin interplay is one of the most generally exploited between biomedical scientists. regardless of its acceptance, avidin-biotin technique suffers from an absence of readability linked to renowned, in your price range, prefabricated kits utilized by many within the box. In Avidin-Biotin Interactions: tools and Applications, leaders in avidin-biotin technique percentage their wisdom with regards to the appliance of the super strong interplay among the protein avidin or its homologues and the diet biotin and a few of its homologues. even though avidin-biotin dependent kits dependent were produced for fairly your time and stay a ordinary method, a number of the chapters during this textual content describe occasions during which the former method failed because of a few issue, and the authors constructed new and higher methods for the avidin-biotin process to be applied. With very good descriptions of laboratory protocols written by way of professional researchers, this quantity is both ideal for the scholar or the pro laboratory scientist. each one bankruptcy in Avidin-Biotin Interactions: equipment and purposes contains not just first-class descriptions of protocols for the lab but in addition the event of researchers who used particular technique, came upon an issue with that method in a brand new atmosphere, and eventually devised how one can reduce or obviate the restrictions of the technology.
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Additional resources for Avidin-Biotin Interactions: Methods and Applications
III (1982) Compositional and structural heterogeneity of avidin glycopeptides. Biochemistry 21, 5334–5341. 9. , Gershoni, J. , Bayer, E. , and Wilchek, M. (1987) Biotin binding to avidin: oligosaccharide side chain not required for ligand association. Biochem. J. 248, 167–171. 10. Green, N. M. (1975) Avidin, in Advances in Protein Chemistry, vol. 29 (Anfinsen, C. , Edsall, J. , and Richards, F. ), Academic Press, New York, pp. 85–133. 11. Chaiet, L. and Wolf, F. J. (1964) The properties of streptavidin, a biotin-binding protein produced by Streptomycetes.
5. 5% NFDM in TBS-T for blocking purposes for 10 min. 6. 5% NFDM. 7. Wash the blots three times with TBS-T for 5 min each. 8. Place the blots face down next to each other on the Storm 840 scanner and scan according to the manufacturer’s instructions. Results for testing the specificity of endogenous biotinylated proteins in rat liver homogenate are presented in Fig. 3. 32 Zwart and Lewis Fig. 3. Avidin competition. The abundance of a particular protein must be considered when deciding which detection method to use.
Eugene, OR, Chapter 4. 17. LaRochelle, W. J. and Froehner, S. C. (1986) Determination of the tissue distributions and relative concentrations of the postsynaptic 43-kDa protein and the acetylcholine receptor in Torpedo. J. Biol. Chem. 261, 5270–5274. 18. Briggs, J. and Panfili, P. R. (1991) Quantitation of DNA and protein impurities in biopharmaceuticals. Anal. Chem. 63, 850–859. 19. Wong, S. S. (1991) Reactive groups of proteins and their modifying agents, in Chemistry of Protein Conjugation and Crosslinking, CRC, Boston, MA, pp.