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  Home > JCE Print > Journal of Chemical Education > Issues > 2007  > September  >
In the Laboratory
Determination of the Subunit Molecular Mass and Composition of Alcohol Dehydrogenase by SDS-PAGE
Barbara T. Nash
Division of Natural Sciences and Veterinary Technology, Mercy College, Dobbs Ferry, NY 10522
Cover
September 2007
Vol. 84 No. 9
p. 1508

Abstract
SDS-PAGE is a simple, rapid technique that has many uses in biochemistry and is readily adaptable to the undergraduate laboratory. It is, however, a technique prone to several types of procedural pitfalls. This article describes the use of SDS-PAGE to determine the subunit molecular mass and composition of yeast alcohol dehydrogenase employing state-of-the-art methods that produce accurate results. Students apply precise amounts of protein to the gel to determine the optimal amount required for accurate measurements. Precision Plus Protein standards are used to produce a standard curve. Students calculate the true subunit molecular mass of the protein using information from the ExPASy protein knowledgebase. They use published data from gel filtration chromatography to determine the molecular mass of the native protein and use this information and their experimental data to determine the subunit stoichiometry of the protein.
Supplement
Student handouts, notes for the instructor, and answers to a postlab problem set are available.
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Citation
Nash, Barbara T. J. Chem. Educ. 2007, 84, 1508.
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Keywords
Bioanalytical Chemistry; Biochemistry; Electrophoresis; Enzymes; Hands-On Learning / Manipulatives; Laboratory Instruction; Noncovalent Interactions; Proteins / Peptides; Upper-Division Undergraduate
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History
Created:
Last Updated:
7/23/2007
8/16/2007
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Experiments, laboratory exercises, lecture demonstrations, and other descriptions of the use of chemicals, apparatus, instruments, computers, and computer interfaces are presented in the Journal of Chemical Education as illustrative of new or improved ideas or concepts in chemistry instruction and are directed at qualified teachers. Although every effort is made to assure and encourage safe practices and safe use of chemicals, the Journal of Chemical Education cannot assume responsibility for uses made of its published materials. Many chemicals are hazardous. Precautions for the safe use of hazardous chemicals and directions for their proper disposal are described in the Material Safety Data Sheets and on the labels. We strongly urge all those planning to use materials from our pages to make choices and to develop procedures for laboratory and classroom safety in accordance with local needs and situations.
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