Firefly luciferase (photinus pyralis)

Description:
Catalyses the oxidation of luciferin to oxyluciferin in an ATP-dependent process generating chemiluminescence at 560 nm (pH 7.8):



Application:
- Assesses the transcriptional activity in cells
- Detects the level of cellular ATP in cell viability assays

Source:
E. coli recombinant enzyme prepared from a thermostable mutant.

Unit Definition:
One light unit produces a biometer peak height equivalent to 0.02 µCi of 14C in PPO/POPOP cocktail. Light units measured in 50 µl assay mixture containing 5 pmol ATP and 7.5 nmol luciferin in Tris-glycine buffer, pH 7.6, at 25°C.

Buffer:
10% Glycerol
50 mM Tris-HCl
1 mM DTT
1 mM EDTA

Storage Conditions:
10 mM Tris-HCl pH 7.8
100 mM NaCl
0.1 mM DTT
0.1 mM EDTA
50% Glycerol

Heat Inactivation:
Active at 42°C for 24 hours (with < 5% loss of activity)

Recommended Storage Condition: -20°C

Reference:
Kricka UJ. (1988) "Clinical and biochemical applications of luciferases and luciferins", Anal Biochem. 175 (1): 14-21