Purification and properties of firefly luciferase
Key findings
The crystalline luciferase purification protocol achieved 9.3-fold enrichment from crude firefly extracts (3,000 units/mg to 28,000 units/mg) through sequential calcium phosphate gel chromatography, stepwise ammonium sulfate fractionation (40-50-60% cuts), and five euglobulin crystallization cycles by dialysis against low ionic strength buffer. Final crystals demonstrated homogeneity by analytical ultracentrifugation and electrophoresis with contaminating pyrophosphatase reduced to zero.