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SRE0045

Sigma-Aldrich

Luciferase from Photinus pyralis (firefly)

recombinant, expressed in E. coli, lyophilized powder, ≥10×1010 units/mg protein

Synonym(s):

Luciferase firefly

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About This Item

CAS Number:
Enzyme Commission number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54

recombinant

expressed in E. coli

Quality Level

form

lyophilized powder

specific activity

≥10×1010 units/mg protein

mol wt

62 kDa

application(s)

diagnostic assay manufacturing

storage temp.

−20°C

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Application

Firefly luciferase is used extensively in molecular and cell biology, in particular for the efficient detection and quantitation of ATP and as a reporter for genetic function.

Biochem/physiol Actions

Firefly luciferase is a 62 kDa protein that catalyzes the production of light. The enzyme requires ATP, molecular oxygen, and the heterocyclic compound, firefly luciferin, to generate light in a two-step process. The light producing reaction is initiated by luciferin activation (adenylation of its carboxylate group) and proceeds in the presence of molecular oxygen to yield a photon of yellow-green light.

Unit Definition

One luciferase enzyme unit will produce one Relative Light Unit (RLU) at 20-25 °C over a 10 second period, measured in 100 μl assay mixture containing 40 pmol ATP and 15 nmol luciferin in Tris-glycine buffer, pH 7.6, using a GloMax 20/20 Luminometer.

Unit Definition Conversion Factor: There are approximately 9000 Relative Light Units (RLU) per one traditional Light Unit that uses a peak height equivalent to 0.02 μCi of 14C in a PPO/POPOP cocktail.

Physical form

Supplied as a lyophilized powder containing HEPES, pH 7.5, NaCl, MgCl2, EDTA, DTT and a carbohydrate stabilizer.
This product is a recombinant luciferase (62 kDa) from Photinus pyralis (American firefly) produced from the luc gene expressed in E. coli.

Preparation Note

To obtain maximal solubility it is important to reconstitute the enzyme at a high salt concentration, such as 1 M Tris buffer with any counter ion at pH 7-8. The enzyme can be prepared at a concentration of up to 5 mg protein/ml. Do not vortex and avoid agitation.
After reconstitution, the enzyme solutions can kept at 4-8 °C for up to 2 days or frozen in working aliquots at -20°C for at least one month. Repeated freezing and thawing is not recommended.

Pictograms

Health hazardCorrosion

Signal Word

Danger

Hazard Statements

Hazard Classifications

Eye Dam. 1 - Resp. Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Pratik Vyas et al.
Journal of the American Chemical Society, 145(15), 8344-8354 (2023-03-24)
Reactions involving the transfer of a phosphoryl (-PO32-) group are fundamental to cellular metabolism. These reactions are catalyzed by enzymes, often large and complex, belonging to the phosphate-binding loop (P-loop) nucleoside triphosphatase (NTPase) superfamily. Due to their critical importance in
S R Ford et al.
Journal of bioluminescence and chemiluminescence, 7(3), 185-193 (1992-07-01)
Commercially available crystalline native and recombinant firefly luciferases were compared. The two types of luciferase had indistinguishable responses to variation in ATP and luciferin concentrations and to omission of reaction components. The time courses of light production, the responses to
Kanika Narula et al.
The Plant journal : for cell and molecular biology, 103(2), 561-583 (2020-03-15)
Pathogen-/microbe-associated molecular patterns (PAMPs/MAMPs) initiate complex defense responses by reorganizing the biomolecular dynamics of the host cellular machinery. The extracellular matrix (ECM) acts as a physical scaffold that prevents recognition and entry of phytopathogens, while guard cells perceive and integrate
Mining Disaggregase Sequence Space to Safely Counter TDP-43, FUS, and I?-Synuclein Proteotoxicity.
Tariq, et al.
Cell Reports, 28, 2080-2095 (2021)
Maura Greiser et al.
eLife, 12 (2023-06-05)
Mitochondrial ATP production in ventricular cardiomyocytes must be continually adjusted to rapidly replenish the ATP consumed by the working heart. Two systems are known to be critical in this regulation: mitochondrial matrix Ca2+ ([Ca2+]m) and blood flow that is tuned

Articles

Firefly luciferase is a sensitive reporter for gene studies due to its absence in mammalian cells or tissues.

Firefly luciferase is a sensitive reporter for gene studies due to its absence in mammalian cells or tissues.

Firefly luciferase is a sensitive reporter for gene studies due to its absence in mammalian cells or tissues.

Firefly luciferase is a sensitive reporter for gene studies due to its absence in mammalian cells or tissues.

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