1) 6 tubes (50 mL) were used
2) Add 50mg/50ml arachidonic acid into each tube and dry out under N₂ stream
3) Add 25ml of cofactor to each tube and add buffer (8109.4 mL) into each tube and vortex
4) Incubate for 10 min at 22 C in shaker bath (preincubation)
5) Add 1865.6 mL of sample ( protein source) into each tube and vortex
6) Incubate for .5 min at 22 C in shaker bath
7) To stop reaction add 1600 mL of 0.1 M HCl into each tube and vortex
8) Extracted with ethyl acetate (3mL was used) three times (1200 rpm 2 min)
9) Extracts were dried under N₂ streams