反应详情

To a 12×75 mm tube containing [1-14C] PGH2 (2 μg, 0.01 μCi or 0.05 μCi) and inhibitor (1 mM final assay concentration, in 10 μl of ethanol) in 225 μl of phosphate buffer (0.1M, pH 7.5), was added to an aliquot of guinea pig lung microsomes (25 μl, containing 400 μg protein) or an aliquot of human platelet microsomes (25 μl, containing ≅130 g protein). The solution was incubated in a 22° water bath for 2 min.; ether (1 ml) and citric acid (25 μMoles) were added, and the tube plunged into an acetone-dry ice bath. The ether extract was transferred to a 1.0 ml Reacti-vial, and the volume reduced under nitrogen to ≅50 μl.

1. concentrationassay concentration
2. additionin 10 μl of ethanol) in 225 μl of phosphate buffer (0.1M, pH 7.5), was added to an aliquot of guinea pig lung microsomes (25 μl, containing 400 μg protein)
3. additionan aliquot of human platelet microsomes (25 μl, containing ≅130 g protein)
4. customwas incubated in a 22°
5. customfor 2 min.
6. customthe tube plunged into an acetone-dry ice bath
7. extractionThe ether extract