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Quantitative determination method of NADH-cytochrome b5 reductase (metmyoglobin-reducing enzyme) in bovine skeletal muscle was developed by combining SDS-PAGE and immunoblotting techniques. Recoveries of the enzyme by this assay ranged from 90.0 to 94.4%. The content of NADH-cytochrome b5 reductase in bovine skeletal muscle was estimated to be 13.8±2.6(9.0-18.2)μg/g tissue (n=19). Also, a quantitative assay for determining cytochrome b5 (an electron transfer mediator for metmyoglobin reduction) was similarly established and cytochrome b5 content in bovine skeletal muscle was estimated to be 59.0±20.9μg/g tissue. The quantitative data of NADH-cytochrome b5 reductase in this study supported our previous hypothesis that the enzyme plays a critical role in preventing metmyoglobin accumulation in muscles. Anim. Sci. Technol. (Jpn.) 68 (1): 29-33, 1997

Quantification of NADH-Cytochrome b5 Reductase (Metmyoglobin-Reducing Enzyme) in Bovine Skeletal Muscle by an Immunoblotting Assay