Figure 7.

(A) Separation of an Orfelia fultoni larvae extract and bioluminescence of luciferin- and luciferase- containing fractions. An extract of 4 frozen Orfelia larvae was prepared by grinding the organisms in liquid N₂, mixing with 0.10 M pH 7.0 ammonium acetate buffer containing 1 mM EDTA. After centrifugation, 75 uL of the supernatant was applied to a Yarra 300 × 4.6 mM SEC-3000 size exclusion column eluted at a flow rate of 1 mL/min on a Thermo Finnegan Surveyor HPLC system with total UV monitoring. The inset shows a photograph taken in the dark of 0.4 mL of Fractions 2 and 3 mixed together in an Eppendorf tube; (B) SDS-PAGE of an Orfelia fultoni larvae extract before and after size exclusion chromatography. The crude larvae extract (middle lane) and luciferase activity-containing concentrated Fraction 2 from the experiment illustrated in panel A were applied and run on a gradient SDS gel.