Reporter gene assays and in vivo imaging

Methodology:
The luc gene coding for firefly luciferase may be attached to another gene of interest. The expressing of this gene may then be estimated from the level of luciferase expression measured as light. This may be done in vitro, i.e. after lysing the cells adding ATP and D-luciferin, or in vivo. In the latter case D-luciferin has to enter the cells. Even whole live animals or plants may be studied, “in vivo imaging”, by injecting D-luciferin into the organism (ATP is already in the cells).

Intended use:

1. In an easy and sensitive way measure the expression of a gene of interest under various conditions by measuring the expression of luciferase.
2. Follow how luminescent cells or viruses spread in e.g. an animal.

Suitable products for in vitro measurements: Luciferase Assay Kit, Luciferase Assay Kit HTS

Suitable products for in vivo measurements: Endotoxin free D-luciferin, potassium salt

Recommended papers:

Lundin, A. (1993) Optimised assay of firefly luciferase with stable light emission. In: Bioluminescence and Chemiluminescence (Szalay, A., Kricka, L. J. and Stanley, P., Eds.), pp. 291-295, John Wiley & Sons, Chichester.

Lundin A. (2014) Optimization of the firefly luciferase reaction for analytical purposes. Adv. Biochem. Engin./Biotechnol. 2014; 145:31-62

El-Sayed, R., Eita, M., Barrefelt, Å., Ye, F., Jain, H., Fares, M., Lundin, A., Crona, M., Abu-Salah, K., Muhammed, M., and Hassan, M. Thermostable Luciferase from Luciola cruciate for Imaging of Carbon Nanotubes and Carbon Nanotubes Carrying Doxorubicin Using in Vivo Imaging System. (2013) Nano Lett. 13, 1393−1398