10411523001

Luciferase

from Photinus pyralis (American firefly)

• Enzyme Commission number: 1.13.12.7 (BRENDA, IUBMB)
• UNSPSC Code: 12352204

Properties

• form: lyophilized (recrystallized twice)
• specific activity: ~8 units/g protein (At 25 °C with 0.1 mM luciferin and 0.7 mM ATP as the substrates (0.1 M Tris-acetate, pH 7.75).)
• packaging: pkg of 1 mg
• manufacturer/tradename: Roche
• optimum pH: 7.5
• shipped in: wet ice

Description

General description

Luciferase is a class of enzymes naturally found in organisms like protists, fungi, insects, bacteria which belong to phylogenic kingdom. It is used as a molecular marker for biological imaging as it specifically cleaves luciferin substrate which results in emitting light.

Luciferase is an enzyme, that produces light. It is present in insect fireflies and in luminous marine and terrestrial microorganisms.[1] Luciferase proteins are available in two forms, such as firefly (Photinus pyralis) and bacterial.[2]

Application

Luciferase has been used as a component in Mix B respiration medium for the determination of the ATP synthetic activity of the H⁺-ATP synthase.[3] It has also been used in thermal aggregation to assay CnoX (YbbN) activity.[4]

Biochem/physiol Actions

Firefly luciferase oxidizes luciferin to oxyluciferin using molecular oxygen and ATP in the reaction and liberates light at 560 nm.[2]

Quality

Contaminants: <1 mU ATPase/mg lyophilizate, and <10 mU nucleoside diphosphatase kinase/mg lyophilizate. The lyophilizate contains neither luciferin nor arsenate.

Sequence

Enzyme consists of two subunits, one of them is active.

Preparation Note

Storage conditions (working solution): -15 to -25 °C
Store the reconstituted solution in aliquots at -15 to -25 °C.
Avoid repeated freezing and thawing.
Information Note: Do not store in plastic vessels.

Reconstitution

Cool the original glass vial (1 mg or 5 mg protein, respectively) to 0 °C in an ice-bath and dissolve the containing luciferase by adding 1 ml or 5 ml, respectively, 0.5 M ice-cold Tris-acetate buffer, pH 7.5, and let it stand for 10 minutes. (The dissolved enzyme is sensitive to shaking and to storage in plastic vessels). This concentrated solution is stable for several
months when stored frozen in portions of approximately 50 μl. Diluted solutions are unstable and should be prepared directly before use and should be kept in an ice-bath.
One microgram luciferase corresponds to approximately 10⁷ arbitrary luciferase units.

Storage and Stability

Store at 2 to 8 °C. (A decrease in activity of approximately 20% may occur within 12 months.)

Other Notes

For life science research only. Not for use in diagnostic procedures.

Safety Information

• Storage Class: 11 - Combustible Solids
• wgk_germany: WGK 1
• flash_point_f: does not flash
• flash_point_c: does not flash