Activity: 1-naphthol sulfotransferase
Incubation conditions
Assay buffer: 0.05 M potassium phosphate pH 7.4, 5 mM MgCl2, 10 mM dithiothreitol
Incubation volume: 200 µl
Stop reagent: acetonitrile (100 µl)
HPLC conditions
Column: Hypersil ODS (5 µm) 250 x 4 mm
Temperature: 30°C
Mobile phase: 5% (v/v) triethylamine (pH 3.0)/methanol; 47/53 (v/v) (isocratic)
Flow rate: 1.0 ml/min
Run time: 8.5 min
Injection volume: 50 µl
Detection: fluorescence λ(ex) = 280 nm, λ(em) = 335 nm
Retention times:
1-naphthylsulfate 3.5 min
1-naphthol 7.3 min