Prev. Article Next Article Table of Contents

Article

A Convenient and Highly Specific Western Blot Experiment for Introductory Biochemistry

Supporting Info

Your current credentials do not allow retrieval of the full text.

Purchase the full-text

PDF/HTML,
figures/images,
references and tables,
(where available)

Donald G. Gerbig Jr.

Biology Department, Kent State University-East Liverpool Campus, East Liverpool, OH 43920

Christopher J. Fenk and Stephanie Y. Grooms

Chemistry Department, Kent State University-East Liverpool Campus, East Liverpool, OH 43920

J. Chem. Educ., 2000, 77 (3), p 373

DOI: 10.1021/ed077p373

Publication Date (Web): March 1, 2000

Abstract

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.