Recombinant Firefly Luciferase (firefly) Protein

Product Discontinued

Recombinant Firefly Luciferase (firefly) Protein Summary

• Description: Recombinant Luciferase is a luciferase expressed from a cloned gene from the North American firefly (Photinus pyralis).
• Preparation Method: E. coli, Photinus pyralis
• Source: E. coli
• Protein/Peptide Type: Full Length Recombinant Protein
• Purity: >95%, by SDS-PAGE

Applications/Dilutions

• Application Notes: The light unit value is the light output generated at room temperature over a 10-second period. Light output is measured using a Turner Luminometer Model# TD-20e. Specific Activity is 2 x 10^10 Light Units/mg protein.
• Theoretical MW: 60.75 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Luciferase from Firefly.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: 25mM Tris-acetate, pH 7.8 (4C), 1mM EDTA, 1mM DTT, 0.2M ammonium sulfate, 15% glycerol, and 30% ethylene glycol.
• Preservative: No Preservative
• Concentration: 12.5 mg/ml
• Purity: >95%, by SDS-PAGE

Alternate Names for Recombinant Firefly Luciferase (firefly) Protein

• firefly luciferase

Background

Luciferase is a generic term for a group of oxidative enzymes used in bioluminescence. Firefly (Photinus pyralis) and bacterial luciferase enzymes are commonly used in assay systems such as cell viability assays, reporter gene assays, and for in vivo imaging. Bacterial luciferases are flavoenzymes composed of two subunits each encoded by the luxA and luxB genes, while the firefly luciferase is a single polypeptide specified by the luc gene (1). Firefly luciferase (theoretical molecular weight: 61 kDa) oxidizes the substrate luciferin to oxyluciferin in a bioluminescent reaction requiring Mg2+ and ATP (2,3). This reaction produces a flash of yellow-green light with an emission peak around 560nm that can be detected by a luminometer (3). Firefly luciferase has become one of the more widely used reporter proteins and is an excellent tool for the study of gene expression, given that the amount of light emitted is directly proportional to luciferase activity (4).

The luciferase assay is fast and sensitive, differentiating itself from the CAT (chloramphenicol acetyltransferase) assay because it does not require a radioactive substrate.

References

1. Eun, H. (1996). Marker/Reporter enzymes. Enzymology Primer for Recombinant DNA Technology, 567-645. doi:10.1016/b978-012243740-3/50011-9

2. McNabb, D. S., Reed, R., & Marciniak, R. A. (2005). Dual luciferase assay system for rapid assessment of gene expression in Saccharomyces cerevisiae. Eukaryotic Cell, 4(9), 1539-1549. doi:10.1128/ec.4.9.1539-1549.2005

3. Fraga, H. (2008). Firefly luminescence: A historical perspective and recent developments. Photochemical & Photobiological Sciences, 7(2), 146-158. doi:10.1039/b719181b

4. Younes, A., Lukyanenko, Y. O., Lyashkov, A. E., Lakatta, E. G., & Sollott, S. J. (2011). A bioluminescence method for direct measurement of phosphodiesterase activity. Analytical Biochemistry, 417(1), 36-40. doi:10.1016/j.ab.2011.05.036

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Peptides and proteins are guaranteed for 3 months from date of receipt.

⚠ WARNING: This product can expose you to chemicals including Ethylene Glycol, which is known to the State of California to cause reproductive toxicity with developmental effects.  For more information go to www.P65Warnings.ca.gov.

Publications for Luciferase (firefly) Full Length Recombinant Protein (NB810-74573)(1)

Publication using NB810-74573

• Wahyudi H, Wang Y, McAlpine SR et al. Dimerization of a heat shock protein 90 inhibitor enhances inhibitory activity. Org Biomol Chem. 2013-12-10 [PMID: 24323090]