MNARAMURA at atlas.niaid.nih.gov
Tue Feb 25 16:39:49 EST 1997
The excitation wavelength of the original GFP from jellyfish (wtGFP)
was not really suitable for 488 nm laser. Also, jellyfish codon usage
was not efficiently translated by mammalian cells. I remember there
was also some discussion on temperature sensitivity. But now there are
a lot of commercially available GFP mutants which were actually
selected for detection by flow cytometry. I have used pGreenLantern-1
from GIBCO BRL and pEGFP-1 from CLONTECH, and both worked very well in
flow. I compared pGreenLantern-1 with wtGFP in the same cell line, and
the difference in the mean fluorescence intensity was 2 to 3 log's.
Mayumi Naramura, M.D.
Lab of Immunology
Twinbrook II, Rm. 125
12441 Parklawn Drive
Rockville, MD 20852
e-mail:mnaramura at atlas.niaid.nih.gov
From: David L. Haviland, Ph.D.
Sent: Tuesday, February 25, 1997 11:29
I was curious if people had been successfully analysed GFP levels
transfected cells using flow?
I once had an investigator try something like this (though I do *not*
if it was GFP, per se) and the experiment was a complete bust! There
something about the excitation/emmision that was not near
there was enough delay that no fluoresence could be detected except
using a flourescent microscope.
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