HPLC Estradiol Disulfate, PF-04859989 Reversibility

0.7 µg of KAT-II was incubated at 37°C for 10 minutes in a 50 µL reaction mixture containing PLP (10 µM, 50 µM, 100 µM, 200 µM), α-ketoglutarate (5 mM), L-KYN (5 mM) in PBS, pH 7.4, and the inhibitor studied (1-2000 µM). Following incubation, the reaction was terminated by adding formic acid (0.8 M) in a 1:1 ratio. 50 µL of the mixture was transferred into a vial with 950 µL of water for HPLC analysis.The production of KYNA was measured using HPLC with UV detection at a wavelength of 330 nM, using a C18 reverse-phase column, and 50% (v/v) water and 50% (v/v) methanol mobile phase. The data was collected in triplicate.