Thursday, November 22, 2012

Applications Thursday – Dual Glow Reporter Gene Assays Benefit from Dual Emission Detection and Longer Bandwidth Filters

A new second wave of dual glow luciferase reporter gene assays are now available from various companies (Pierce, Promega, Life Technologies). Other than Firefly and Renilla, there are now Gaussia, Cypridina, Green Renilla, and Red Firefly luciferases that are available and that can be multiplexed together, or even with fluorescent signals.

One main difference between these new dual glow reporter assays  and earlier dual glow assays like DLR™, is that the two different luminescent signals can be differentiated at different wavelengths, rather than at different time points. This enables higher throughput because there is no need for injection and no need for a minimal measurement time, the only limitation is in the instrumentation.

Larger bandwidth filters can capture more signal. 
Two features on instruments that will benefit these dual glow assays are Dual Emission Detection, and Longer Bandwidth Filters. Since two emission wavelengths need to be measured per well, microplate readers with Dual Emission Detection, like the POLARstar Omega and PHERAstar FS, will measure these assays in half the time compared to instruments that do not have Dual Emission Detection. Dual Emission Detection also helps to correct with read-to-read variations that can occur with reading the plate twice.

In addition, since these luminescent signals are not as bright as fluorescent signals, it benefits from collecting a larger area of the emission signal at each wavelength. Thus microplate readers with the ability to measure broad bandwidths (50-100 nm), like the POLARstar Omega and PHERAstar FS because they use longer bandwidth filters, will be more sensitive in these assays.

Learn more about it here, Wavelength Based Dual Glow Reporter Genes.