Cypex CYP2E1 QC Assays
buffer: 0.1 M potassium phosphate pH 7.4, 5 mM MgCl2
volume: 200 µl
Stop reagent: 1 M hydrochloric acid (25 µl)
Internal standard: zoxazolamine (10 µl of 100 µg/ml)
Column: Hypersil BDS-C18 (3 µm) 100 x 4 mm
Mobile phase: 0.05% orthophosphoric acid/acetonitrile 81/19 (v/v)
Flow rate: 1.0 ml/min
Run time: 10.8 min
volume: 50 µl
Detection: UV, l
= 295 nm
of phosphate concentration on chlorzoxazone turnover by CYP2E1R Bactosomes
turnover by CYP2E1 is sensitive to the concentration of phosphate in the incubation.
assay was carried out at 500 µM chlorzoxazone at a P450 concentration of
10 pmol per 200 µl.
The length of time for which product formation is
linear is also affected by phosphate concentration.
under licence from BTG International Ltd (AU730155, EP0914446, US6566108 and other
United States Patent Nos. 5,420,027 or 5,240,831, Canada
Patent No. 2100245 and other patents pending.