| Assay Method Information | |
| | VKOR Activity Assay |
| Description: | Briefly, standard reactions were performed in 200 mM Hepes buffer, pH 7.4, containing 150 mM KCl, 1 mM dithiothreitol, 0.25 to 2 g liter<sub>-1</sub> of total proteins containing membrane VKORC1. The reaction was started by the addition of vit K1>O solution in 1% Triton X-100 and incubated at 37 °C for 30 min. In these conditions, the reaction was linear according to the time of incubation and the quantity of incubated proteins. After incubation at 37 °C for 30 min, the reaction was stopped by adding 4 ml of iced 1:1 isopropyl alcohol/hexane solution. After centrifugation at 5000 x g for 10 min, the hexane layer was removed and driedunder nitrogen. The dry residue was immediately dissolved in 0.2 ml of isopropyl alcohol, and the reaction product was analyzed by liquid chromatography-mass spectrometry. |
| Affinity data for this assay | |
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