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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Microplate Assay Measurement of Cytochrome P450-Carbon Monoxide Complexes

  • Choi, Suk-Jung (Department of Chemistry and East Coastal Marine Bioresources Research Center, Kangnung National University) ;
  • Kim, Mi-Ra (Department of Chemistry and East Coastal Marine Bioresources Research Center, Kangnung National University) ;
  • Kim, Sung-Il (Department of Chemistry and East Coastal Marine Bioresources Research Center, Kangnung National University) ;
  • Jeon, Joong-Kyun (Faculty of Marine Bioscience and Technology, Kangnung National University)
  • Received : 2003.01.03
  • Accepted : 2003.01.30
  • Published : 2003.05.31
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Abstract

Cytochrome P450 in microsomes can be quantitated using the characteristic 450 nm absorption peak of the CO adduct of reduced cytochrome P450. We developed a simple microplate assay method that is superior to previous methods. Our method is less laborious, suitable for analyzing many samples, and less sensitive to sample aggregation. Microsome samples in microplate wells were incubated in a CO chamber rather than bubbled with CO gas, and then reduced with sodium hydrosulfite solution. This modification allowed a reliable and reproducible assay by effectively eliminating variations between estimations.

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References

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