Cypex CYP4F2 QC
Activity: Leukotriene B4
buffer: 0.1 M potassium phosphate pH 7.4, 5 mM MgCl2
Incubation volume: 50 µl
Stop reagent: acetonitrile/acetic acid, 98/2 (v/v) (25 µl)
Internal standard: not used
Column: HyperClone ODS (5 µm) 250
x 4 mm
Mobile phase: 100 mM ammonium acetate (pH 5.6)/water/methanol 20/35/45
(% v/v/v) initially (gradient separation)
Flow rate: 1.0 ml/min
Run time: 15.5 min
Injection volume: 50 µl
Detection: UV (DAD), l
= 271 nm
of phosphate concentration on leukotriene B4
turnover by CYP4F2BLR
B4 turnover by CYP4F2 is very sensitive to the
phosphate in the incubation.
assay was carried out at 100 µM leukotriene B4
at a P450
concentration of 2.5 pmol per 50 µl.
under licence from BTG International Ltd (AU730155, EP0914446,
US6566108 and other patents pending).
United States Patent Nos. 5,420,027 or 5,240,831, Canada Patent No.
2100245 and other patents pending.