GFP

Mayumi Naramura 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.
NIH/NIAID
Lab of Immunology
Twinbrook II, Rm. 125
12441 Parklawn Drive
Rockville, MD 20852
Phone:  301-402-4595
FAX:      301-594-2522
e-mail:mnaramura at atlas.niaid.nih.gov

----------
From: 	David L. Haviland, Ph.D.
Sent: 	Tuesday, February 25, 1997 11:29
To: cyto-inbox
Subject: 	GFP


Hi:

I was curious if people had been successfully analysed GFP levels 
within
transfected cells using flow?
I once had an investigator try something like this (though I do *not* 
know
if it was GFP, per se) and the experiment was a complete bust!  There 
was
something about the excitation/emmision that was not near 
instantaneous and
there was enough delay that no fluoresence could be detected except 
when
using a flourescent microscope.

David






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