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  Home > JCE Print > Journal of Chemical Education > Issues > 2000  > March  >
In the Laboratory
A Convenient and Highly Specific Western Blot Experiment for Introductory Biochemistry
Christopher J. Fenk and Stephanie Y. Grooms
Chemistry Department, Kent State University-East Liverpool Campus, East Liverpool, OH 43920

Donald G. Gerbig Jr.
Biology Department, Kent State University-East Liverpool Campus, East Liverpool, OH 43920

Cover
March 2000
Vol. 77 No. 3
p. 373

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.
Supplement
Objectives, background information, experimental notes, Post Lab questions, and figures are included.
*  Contents
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More Information
*  Citation
Fenk, Christopher J.; Grooms, Stephanie Y.; Gerbig, Donald G., Jr. J. Chem. Educ. 2000 77 373.
*  Keywords
Biochemistry; Laboratory Instruction; Enzymes; Proteins / Peptides; Separation Science; Intermolecular Forces
*  History
Created:
Last Updated:
February 14, 2000
August 31, 2005
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