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L9506 Sigma

Luciferase from Photinus pyralis (firefly)

BioUltra, ≥98% (SDS-PAGE), lyophilized powder, 15-30×106 light units/mg protein

Synonym: Luciferase firefly

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Properties

Related Categories 1.13.x.x Acting on single donors with incorporation of molecular oxygen, 1.x.x.x Oxidoreductases, Analytical and Industrial Enzymes, Application Index, Biochemicals and Reagents,
Quality Level   PREMIUM
product line   BioUltra
assay   ≥98% (SDS-PAGE)
form   lyophilized powder
composition   Protein, 10-35% E1%/280
measuring range   ≤1 femtomole ATP sensitivity, (using 0.2 μg of luciferase and suitably sensitive liquid scintillation counters or luminometers)
Featured Industry   Diagnostic Assay Manufacturing
foreign activity   ATPase ≤5 nanomolar units/mg protein
  Nucleoside diphosphokinase ≤20 nanomolar units/mg protein
shipped in   wet ice
storage temp.   −20°C

Description

Analysis Note

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.

General description

Firefly luciferase is an enzyme that catalyzes production of light from luciferin in the presence of Mg2+-ATP and oxygen. The reaction of this enzyme with luciferin, ATP, and O2 results in the emission of light.

Other Notes

Arsenate free.

Firefly luciferase is also available in economical recombinant forms (Product Numbers SRE0045 and L9420)

Packaging

Sold on the basis of protein content

Preparation Note

Chromatographically prepared and crystallized.

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.

Physical form

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

Application

This enzyme has wide range of applications in biotechnology and development of biosensors. Luciferase can be used to detect trace amounts of ATP and is 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. This enzyme has been used in a study to identify the different characteristics of reporter genes in whole-cell bacterial sensors. Luciferase from Photinus pyralis has also been used in a study to develop a novel bioluminogenic assay for α-chymotrypsin.

Biochem/physiol Actions

Luciferase activity can be inhibited by general anesthetics including isoflurane and ketamine/medetomidine thereby affecting the sensitivity of bioluminescence imaging.

Price and Availability


Amplified Detection

All labs need water

Biomedical Applications
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3

Documents

Certificate of Analysis

Certificate of Origin

Protocols & Articles

Articles

ABC Transporters and Cancer Drug Resistance

Chemotherapy is the treatment of choice against many types of cancer. However, over time chemotherapeutic drugs can become less effective due to the development of resistance that involves a group of...
BioFiles 2007, 2.4, 7.
Keywords: Absorption, Cancer, Clinical, Cloning, Combinatorial synthesis, Diseases, Enzymology, Gene expression, Genetic, Immunology, Pharmaceutical, Reductions, Search, Transfection, Vectors

TokeOni: Properties and Applications

Bioluminescence imaging (BLI) systems allow for high-sensitive and noninvasive monitoring of cell proliferation, activity of signaling pathways and protein-protein interactions in living tissues.1-3 ...
Takahiro Kuchimaru and Shinae Kizaka-Kondoh
Graduate School of Bioengineering & Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Keywords: Cancer, Cell proliferation, Infrared spectroscopy

Protocols

Enzymatic Assay and ATP Sensitivity Test of Luciferase

To standardize a procedure to determine the enzymatic activity of Luciferase and/or to determine the ATP detection limit of Luciferase.
Keywords: Biochemistry, Enzyme activity, Enzymology, Size-exclusion chromatography

Related Content

Enzymes & Proteins

Application Index | Enzyme Index | Substrate Index | Inhibitor Index | Cofactor Index | Lectin Index
Keywords: Cell signaling, Diagnostic, Drug discovery, Molecular biology

Peer-Reviewed Papers
15

References

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L9420 Luciferase from Photinus pyralis (firefly), recombinant, expressed in E. coli, buffered aqueous solution, ≥10×1010 units/mg protein

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SRE0045 Luciferase from Photinus pyralis (firefly), recombinant, expressed in E. coli, lyophilized powder, ≥1×1011 units/mg protein

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L6882 D-Luciferin sodium salt, lyophilized powder, synthetic

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