Re: GFP transfection

From: ckuszyns@UNMC.EDU
Date: Mon Feb 18 2002 - 11:05:34 EST


It has been our experience that some of the transfection methode increase
the autofluorescence of cells.  We routinely analyze eGFP transduced EC
cells which grwo as adherent cells and then are removed with trypsin.  We
recently compared the efficiency of transfection with different compounds
compared to standard CaPO4 methods.  We found that expression was enhanced
significantly using Lipofectamine  (PLUS) and Lipofectamine 2000.  The
increases were in both the percent of cells expressing the fluorescent
protein and in the intensity of fluoresence per cell.

We use a FACS Calibur and have not changed anything to detect this protein.

Charles A. Kuszynski, Ph.D.
Assistant Professor/Director
Cell Analysis Facility
University of Nebraska Medical Center
986495 Nebraska Medical Center
Omaha, NE 68198-6495
402 559-6267 voice
402 559-4077 fax
ckuszyns@unmc.edu



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