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

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  • ARIHARA Keizo
    School of Veterinary Medicine and Animal Sciences, Kitasato University
  • ITOH Makoto
    School of Veterinary Medicine and Animal Sciences, Kitasato University
  • KONDO Yo
    School of Veterinary Medicine and Animal Sciences, Kitasato University

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  • 牛骨格筋におけるNADH-チトクロムb5還元酵素(メトミオグロビン還元酵素)の免疫プロッティング法による定量
  • Quantification of NADH Cytochrome b5 Re

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Abstract

Quantitative determination method of NADH-cytochrome b5 reductase (me- tmyoglobin-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 cyto- chrome 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.

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