RE: GFP

Mayumi Naramura (MNARAMURA@atlas.niaid.nih.gov)
Tue, 25 Feb 1997 16:39:49 -0500

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@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


Home Page Table of Contents Sponsors E-Mail Archive Web Sites

CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone: (765)-494-0757; FAX(765) 494-0517; Web http://www.cyto.purdue.edu , EMAIL cdrom3@flowcyt.cyto.purdue.edu