Cypex SULT2A1 QC Assay

Activity: 17beta-estradiol sulfotransferase (3-sulfate formation)

Incubation conditions
Assay buffer: 0.05 M potassium phosphate pH 7.4, 5 mM MgCl₂, 10 mM dithiothreitol
Incubation volume: 200 µl
Stop reagent: 1 M hydrochloric acid (10 µl)

HPLC conditions
Column: HyperClone ODS (5 µm) 250 x 4 mm
Temperature: 40°C
Mobile phase: (isocratic)
    41% Water
    45% Acetonitrile
    14% 50 mM ammonium acetate/50 mM tetrabutylammonium (pH 3.6)
Flow rate: 1.0 ml/min
Run time: 9.5 min
Injection volume: 100 µl
Detection: fluorescence l(ex) = 211 nm, l(em) = 312 nm
Retention times:
       17beta-estradiol 17-sulfate    3.9 min
     17beta-estradiol 3-sulfate      4.4 min
     17beta-estradiol                   5.5 min

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United States Patent Nos. 4,952,496, 5,693,489 and 5,869,320.