Microplate assay measurement of cytochrome p450-carbon monoxide complexes.
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.
Choi
Suk-Jung
SJ
Department of Chemistry, Kangnung National University, Gangneung 210-702, Korea. sjchoi@kangnung.ac.kr
Kim
Mira
M
Kim
Sung-Il
SI
Jeon
Joong-Kyun
JK
D000818:Animals; D002248:Carbon Monoxide / Q000737:chemistry; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis*; D008862:Microsomes, Liver / Q000201:enzymology; D051381:Rats; D015203:Reproducibility of Results; D013053:Spectrophotometry / Q000379:methods*; D013440:Sulfides / Q000737:chemistry; D013997:Time Factors
D013440:Sulfides; D002248:Carbon Monoxide; D003577:Cytochrome P-450 Enzyme System; C025451:sodium bisulfide