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Gaussia Luciferase Data Sheet

Catalog No. 0308

Product Description

Gaussia luciferase is a luminescent protein with quantum yields higher than the luciferases from Renilla sp. and firefly. The enzyme catalyzes the oxidation decarboxylation of coelenterazine to give light (λmax = 485nm) and coelenteramide. ATP is not needed for activity. Gaussia luciferase is salt tolerant and retains it activity in the presence of non-ionic detergents, cholate, and deoxycholate. In addition, the enzyme is thermal tolerant and can retain activity at temperatures up to 60C. The protein is also resistant to irreversible denaturation. Enzymatic activity can be recovered after treatment with 7M guanidine HCl or 8M urea in the presence of detergent.

Product Specifications
Protein 1mg (0308-1), 250g (0308-2)
Long-Term Storage 2 years at - 80C
Aliquot to avoid repeated freezing and thawing.
Short-Term Storage 1 month at 4C
Formulations In: 10mM Na2HPO4, 140mM NaCl, 2 mM KH2PO4, 3mM KCl, 10% glycerol, pH 7.6
Molecular Weight 22kDa by SDS- PAGE

Instructions for Use

1. The amount of luciferase used for a given assay should be empirically determined by titering the enzyme. The amount of luciferase that gives the highest signal: noise ratio should be selected.
2. Prepare a colenterazine stock by dissolving the powder in 1mL of acidified methanol (50L concentrated HCl to 10mL of anhydrous methanol) to give a 200M solution. Aliquot and store at -80C for up to 4 weeks. A working solution of coelenterazine is prepared by diluting the stock solution to 10M in 18Mohm water.
3. For performing luciferase assays, dilute the enzyme in 50mM Tris-Cl, 0.5M NaCl, pH 7.8 to the desired RFU per assay. Add coelenterazine to 2.5M and immediately measure the light production at 485nm with a 5-10 second integration.

Effects of salt concentration and pH on light emission of Gaussia luciferase

The graph on the top shows the emission spectrum for Gaussia luciferase (λemax= 485nm). The lower graph shows the effects of salt concentration and pH on light emission of Gaussia luciferase.

Material Safety Data

FOR RESEARCH USE ONLY. NOT INTENDED OR APPROVED FOR HUMAN, DIAGNOSTICS OR VETERINARY USE. Do not ingest, swallow or inhale. Do not get in eyes, on skin, or on clothing. Wash thoroughly after handling. For complete safety information see full Material Safety Data Sheet.

References

1. Verhaegen, M. and Christopoulos, T. K. 2002. Recombinant Gaussia Luciferase. Overexpression, Purifi cation, and Analytical Application of a Bioluminescent Reporter for DNA Hybridization. Anal. Chem. 74: 4378-4385.