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Updated: Aug 8, 2026

Preparation of Rat Skeletal Muscle Homogenates for Nitrate and Nitrite Measurements
Published on: July 29, 2021
Use of protein in extraction and stabilization of nitrate reductase
L E Schrader1, D A Cataldo, D M Peterson
1Department of Agronomy and United States Department of Agriculture, Agricultural Research Service, University of Wisconsin-Madison, Madison, Wisconsin 53706.
Abstract:
The in vitro instability of nitrate reductase (EC 1.6.6.1) activity from leaves of several species of higher plants was investigated. Decay of activity was exponential with time, suggesting that an enzyme-catalyzed reaction was involved. The rate of decay of nitrate reductase activity increased as leaf age increased in all species studied. Activity was relatively stable in certain genotypes of Zea mays L., but extremely unstable in others. In all genotypes of Avena sativa L. and Nicotiana tabacum L. studied, nitrate reductase was unstable. Addition of 3% (w/v) bovine serum albumin or casein to extraction media prevented or retarded the decay of nitrate reductase activity for several hours. In addition, the presence of bovine serum albumin or casein in the enzyme homogenate markedly increased nitrate reductase activity (up to 15-fold), especially in older leaf tissue.
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