| Assay Method Information | |
| | In Vitro HO Activity Assay |
| Description: | HO activity in rat spleen (HO-1) and brain (HO-2) microsomal fractions was determined by the quantitation of CO formed from the degradation of methemalbumin. Incubations for HO activity analysis were done under conditions for which the rate of CO formation was linear with respect to time and microsomal protein concentration. Briefly, reaction mixtures consisting of 100 mM phosphate buffer (pH 7.4), 50 uM methemalbumin, and 1 mg/mL protein were pre-incubated with the inhibitors at final concentrations ranging from 0.1 to 100 uM for 10 min at 37 deg C. Reactions were initiated by adding NADPH at a final concentration of 1 mM and incubations were performed for an additional 15 min at 37 deg C. Reactions were stopped by instantly freezing the reaction mixture on dry ice, and CO formation was monitored by gas chromatography. |
| Affinity data for this assay | |
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