Recombinant Luciferase is a luciferase expressed from a cloned gene from the North American firefly (Photinus pyralis) that provides the reliability and dependability needed for performing research or producing kits using bioluminescence reagents to detect ATP or luciferin substrates. A recombinant source eliminates the possibility of seasonal and regional variability that may be found in luciferase purified from natural sources.
Firefly luciferase is a 62kD 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. Luciferase catalyzes the oxidation of reduced flavin mononucleotide (FMNH2) and a long chain aldehyde by molecular oxygen to yield FMN, the corresponding acid, H2O and light.
Source:Recombinant (E. coli, Photinus pyralis)
EC No:1.14.14.3
Molecular Weight:62kD
Purity:≥98% by SDS PAGE
Protein Concentration:10-35% (E0.1% t 280nm=0.75)
Specific Activity:≥20 Billion units/mg protein
Unit Definition: One luciferase enzyme unit will produce one Relative Light Unit (RLU) at 20-25°C over a 10sec period, measured in 100ul assay mixture containing 40pmol ATP and 15nmol luciferin in Tris-glycine buffer, pH 7.6, using a GloMax 20/20 Luminometer.
Preparation Note:To obtain maximal solubility it is important to reconstitute the enzyme at a high salt concentration, such as 1M Tris buffer with any counter ion at pH 7-8. The enzyme can be prepared at a concentration of up to 5mg 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.
Storage and Stability:For long-term storage, aliquot to avoid repeated freezing and thawing and freeze at -20°C. Aliquots remain stable at 95-97% activity through four freeze/thaw cycles. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：1mg
Source Antigen：Recombinant (E. coli, Photinus pyralis)
Grade：Molecular Biology Grade
Purity：≥98% by SDS-PAGE
Form：Supplied as a lyophilized powder containing HEPES, pH 7.5, sodium chloride,, MgCl2, EDTA, DTT and a carbohydrate stabilizer.
