Live cell substrate is a uniquely engineered coelenterazine derivative that can generate up to a five-fold brighter signal than wild-type coelenterazine in the presence of Renilla luciferase
A substrate specifically designed for use with live cells is now available from Promega.
The ViviRen live cell substrate is described as a uniquely engineered coelenterazine derivative that can generate up to a five-fold brighter signal than wild-type coelenterazine in the presence of Renilla luciferase.
In addition, when used with live cells, it can produce up to a 100-fold higher signal-to-noise ratio, says Promega.
The key to the high sensitivity is the reduction in autoluminescence occurring as a result of enzyme-independent substrate breakdown in aqueous environments (for example, culture medium).
ViviRen live cell substrate has been engineered to protect the oxygenation site to prevent autoluminescence and maintain a low background.
Once transported into cells, intracellular esterases cleave the protecting group to produce coelenterazine, the substrate for Renilla luciferase.
ViviRen will therefore generate luminescence only in live cells, not in dead or lysed cells.
Consequently, it can be multiplexed with assays that lyse cells with minimal signal carryover.
ViviRen can be used in a variety of cell analysis techniques including reporter gene assays and RNA interference.
It can also be used in bioluminescence resonance energy transfer (Bret) as the substrate permits real-time measurement in multiwell plates.
As the ViviRen is diluted directly into the cell culture growth medium, depending on how the dilutions are performed, the volume per well may increase by as little as 0.1% with the addition of this substrate.
