Hydrochloric Acid → Thromboxane A2

Participants

Reaction Steps

1 ml of a buffer (20 MM Tris-HCl buffer, 1 mM EDTA, pH 7.5) containing human blood platelet microsomes (50 μg protein/ml) and the test compound (final concentration 10−7M) was agitated, then incubated at 0° C. for 30 minutes. To this was added prostaglandin H2 (100 ng/2 μl). This was incubated at 23° C. for 3 minutes to cause a reaction. Next, 1M hydrochloric acid was added to make the solution acidic and stop the reaction, then this was neutralized by 1M Tris-Base and centrifuged at 3000 rpm for 20 minutes. The amount of the TxB2 in the supernatant was measured by the EIA method (Cayman Co. kit).

Provenance & Evidence