GFP in HeLa cells

Beat Ludin ludin at FMI.CH
Mon Apr 28 02:03:52 EST 1997


Joe Binder wrote:

>I have put GFP (ser65thr and ile167thr) into Mengo virus and am using 
>fluorescent microscopy to visualize infected cells.  I have seen low 
>expression levels.  Any advice on preparing/visualizing slides to see 
>the protein better?

1. Using a low-riboflavin medium or a blanaced salt solution should lower 
the background. 
2. Don't fix the cells to get the highest fluorescence output.
3. If fixation is needed, paraformaldehye seems to be the least damaging. 
Readjust the pH to 6.5 or higher.

BTW, I'm not aware of a S65T/I167T mutant, what is its spectrum? Can you 
give me a reference?

Beat


-----------------------------------------------------
Dr. Beat Ludin, FMI, PO Box 2543, 4002 Basel, Switzerland
Tel. +41 61 697 6697 / FAX +41 61 697 3976
Internet:ludin at fmi.ch / Compuserve:100102,1527




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