Activity: Kynuramine deaminase
Incubation conditions
Assay buffer: 90 mM potassium phosphate, pH 7.4
Incubation volume: 200 µl
Stop reagent: 1 M hydrochloric acid (25 µl)
HPLC conditions
Column: HyperClone BDS-C18 (3 µm) 100 x 4.6 mm
Temperature: 30°C
Mobile phase: 10 mM perchloric acid/water/acetonitrile/methanol 50/33/14/3 (v/v/v/v) (isocratic)
Flow rate: 1.0 ml/min
Run time: 5.0 min
Injection volume: 20 µl
Detection: Fluorescence, λ(ex) = 310 nm, λ(em) = 380 nm
Retention times:
4-hydroxyquinoline 2.4 min
[Kynuramine does not fluoresce under these conditions]