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Luciferase vs GFP reporter assays

Bertrand Collet bercollet at netscape.net
Sat Dec 13 11:16:04 EST 2003


Hello,

Luciferase is a great reporter asssay because it is very sensitive. The
assay is dead easy (basically lyse a pellet of transfected cell with
luciferase substrate and measure the light). However
it is destuctive and require a luminometer. For promoter analyses luciferase
is the best system in my opinion. The dual system with Firefly and Renilla
luciferases allow you to correct data
for variation in transfection efficiencies.

GFP is great because it does not require any specific substrate, but you
need either a fluorometer or a microscope equiped for fluoresecence to
visualise GFP expression in situ. Great because non destructive i.e.
visualisation of expression directly on living cells. I use GFP as a marker
to
establish stable cell lines (easy way to check expression for selection of
positive clones, possibility of use in FACS). For promoter analysis I never
tried but the sensitivity will unlikely surpass luciferase. Also many cell
lines have some auto fluorescence that may interfere with the assay:
problematic when sensitivity and accuracy are needed (promoter analyses).

Hope this helps,

Bertrand





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