Luciferase Firefly, Active
Luciferase-like monooxygenase, LUC, EC 1.13.12.7.
Greater than 95.0% as determined by SDS-PAGE.
Description
Luciferase produced in E.Coli is a single, non-glycosylated polypeptide chain containing 335 amino acids (1-311 a.a) and having a molecular mass of 38.5kDa.
Luciferase is fused to a 24 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.
Product Specs
Luciferase, a type of oxidative enzyme involved in bioluminescence, differs from photoproteins. It catalyzes a reaction involving luciferin, Mg2+, and ATP, resulting in the emission of green light at a wavelength of 562 nm. Firefly luciferase is widely employed as a reporter system for investigating gene expression, function, and in drug discovery screening.
This E. coli-derived Luciferase is a single, non-glycosylated polypeptide consisting of 335 amino acids (with the active enzyme encompassing residues 1-311) and possessing a molecular weight of 38.5kDa. It features a 24 amino acid His-tag at the N-terminus and is purified using proprietary chromatographic methods.
The Luciferase solution (0.5mg/ml) is supplied in a buffer containing 20mM Tris-HCl (pH 8.0), 1mM DTT, and 10% glycerol.
Purity exceeds 95.0% as assessed by SDS-PAGE analysis.
The specific activity is greater than 1x109 light units per milligram of protein. One unit of luciferase enzyme activity is defined as the amount required to produce one Relative Light Unit (RLU) at a pH of 7.5 and a temperature of 25°C.
Luciferase-like monooxygenase, LUC, EC 1.13.12.7.
MGSSHHHHHH SSGLVPRGSH MGSHMTSKVY DPEQRKRMIT GPQWWARCKQ MNVLDSFINY YDSEKHAENA VIFLHGNAAS SYLWRHVVPH IEPVARCIIP DLIGMGKSGK SGNGSYRLLD HYKYLTAWFE LLNLPKKIIF VGHDWGACLA FHYSYEHQDK IKAIVHAESV VDVIESWDEW PDIEEDIALI KSEEGEKMVL ENNFFVETML PSKIMRKLEP EEFAAYLEPF KEKGEVRRPT LSWPREIPLV KGGKPDVVQI VRNYNAYLRA SDDLPKMFIE SDPGFFSNAI VEGAKKFPNT EFVKVKGLHF SQEDAPDEMG KYIKSFVERV LKNEQ.
Product Science Overview
Recombinant firefly luciferase is typically expressed in Escherichia coli and purified to a high degree of purity, often exceeding 90% . The enzyme consists of a full-length protein ranging from 1 to 550 amino acids . It belongs to the ATP-dependent AMP-binding enzyme family and is characterized by its ability to catalyze the oxidation of luciferin, resulting in light emission .
The bioluminescent reaction catalyzed by firefly luciferase involves several key components:
- Luciferin: The substrate that undergoes oxidation.
- ATP: Provides the necessary energy for the reaction.
- Mg²⁺: Acts as a cofactor to stabilize the enzyme-substrate complex.
The reaction proceeds as follows:
- Luciferin is activated by ATP to form luciferyl adenylate.
- The luciferyl adenylate is then oxidized by molecular oxygen, producing oxyluciferin and emitting light.
This light emission is what gives fireflies their characteristic glow and is harnessed in various scientific applications .
Firefly luciferase is extensively used as a reporter gene in molecular biology. Its applications include:
- Gene Expression Studies: By linking the luciferase gene to a promoter of interest, researchers can measure the activity of the promoter based on the light emitted.
- Drug Screening: The enzyme’s bioluminescent properties allow for high-throughput screening of pharmaceutical compounds.
- Cellular Imaging: Luciferase can be used to visualize cellular processes in real-time.
The recombinant production of firefly luciferase involves cloning the luciferase gene into an expression vector, which is then introduced into E. coli cells. The bacteria express the luciferase protein, which is subsequently purified using chromatography techniques . This recombinant form retains the enzyme’s bioluminescent properties and is used in various research applications.
