Luciferase from Photinus pyralis (firefly) 

Note: Prior to 1991, a unit of firefly luciferase activity was defined as that amount which will produce 1.0 nanomole of pyrophosphate per minute at pH 7.7, 25 °C, using a system containing 0.6 mM ATP and 0.1 mM D-luciferin. The former nanomolar unit is equivalent to approximately 1.3 x 10⁶ light units.

Two contaminant, ATP-consuming activities are assayed for in this product, ATPase and nucleoside diphosphokinase. These impurities are found to be less than 5 nanomolar units/mg protein and less than 20 nanomolar units/mg protein, respectively.

Arsenate free.

Sold on the basis of protein content

Chromatographically prepared and crystallized.

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.

Lyophilized powder approximately 20% protein; balance is primarily NaCl, HEPES buffer salts, and carbohydrate.

The reaction of this enzyme with luciferin, ATP, and O₂ results in the emission of light. Luciferase can be used to detect trace amounts of ATP. Firefly luciferase is also one of the most commonly utilized reporter genes for the study of gene expression. The bioluminescent reaction catalyzed by luciferase is one of the most sensitive analytical tools for measuring gene expression. Less than or equal to one femtomole of ATP can be detected using 0.2 μg of luciferase.

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lyophilized powder

mol wt 120 kDa (two non-identical subunits, each containing four free thiol groups, one of which is necessary for activity)

Protein, ~20% E1%/280

≤1 femtomole ATP sensitivity, (using 0.2 μg of luciferase and suitably sensitive liquid scintillation counters or luminometers)

ATPase <5 nanomolar units/mg protein

Nucleoside diphosphokinase <20 nanomolar units/mg protein

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−20°C

Stanley, P.E., A review of bioluminescent ATP techniques in rapid microbiology. J. Biolumin. Chemilumin. 4, 375, (1989)

Rajgopal, S., and Vijayalakshmi, M.A. Enzyme Microb. Technol. 6, 482-489, (1984)

Chappelle, E.W., et al., Determination of bacterial content in fluids,. Meth. Enzymol. 57, 65-72, (1978)

Kricka, L.J., Clinical and biochemical applications of luciferases and luciferins. Anal. Biochem. 175, 14-21, (1988)

L6882, D-Luciferin sodium salt