Human SULT1A1*2

SULT1A1 QC Assays

Activity: 4-nitrophenol 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% triethylamine (pH to 3 with H3PO4)/acetonitrile 66/34 (v/v) (isocratic)
Flow rate: 1.0 ml/min
Run time: 10.0 min
Injection volume: 100 µl
Detection: UV, λ = 280 nm

Retention times:
4-nitrophenylsulfate 4.2 min
4-nitrophenol 5.2 min

Activity: 4-methylumbelliferone 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: 50 mM ammonium acetate, 50 mM tetrabutylammonium (pH 3.6)/
acetonitrile; 54/46 (v/v) (isocratic)
Flow rate: 1.0 ml/min
Run time: 7.0 min
Injection volume: 100 µl
Detection: fluorescence λ(ex) = 315 nm, λ(em) = 375 nm

Retention times:
4-methylumbelliferone 3.3 min
4-methylumbelliferylsulfate 4.3 min