A Convenient and Highly Specific Western Blot Experiment for Introductory Biochemistry
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[1] Kent State University, East Liverpool
Kent State University, East Liverpool
City of East Liverpool, Estados Unidos
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- Localización: Journal of chemical education, ISSN 0021-9584, Vol. 77, Nº 3 (March), 2000, págs. 373-373
- Idioma: inglés
- Texto completo no disponible (Saber más ...)
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Resumen
An experiment is described for use in undergraduate biochemistry laboratories. It is designed to take advantage of recent technological improvements in Western blot procedures and may be conveniently conducted in two consecutive 3-hour laboratory periods using readily available reagents. A commercially available mixture of proteins, SDS-PAGE standards, is employed. This eliminates the need to optimize protein concentration for gel loading and immunological detection. In terms of laboratory safety, only proteins from plants and nonprimate animals are used, thus eliminating any concerns related to employing reagents derived from humans or other primates. Use of precast polyacrylamide gels eliminates the hazards associated with handling acrylamide. A chemically inert blotting membrane is employed in place of the more generally used and highly flammable nitrocellulose membrane. These improvements allow for a safe, rapid, and highly selective laboratory protocol that may be easily incorporated into the undergraduate laboratory.
