CAT cell-per-cell detection

Dominique Vercammen nick at lmb1.rug.ac.be
Wed May 11 12:02:09 EST 1994


Dear reader,

	As an essential part of my PhD investigation, I am using chloramphenicol 
acetyltransferase (CAT) as a reporter gene. As I am using it in fusion with 
the gene of my interest, and expressing it in a transient way, I would like to 
make a distinction between cells being effectively transfected, and those who 
are not.

	I know that antibodies are available against CAT, which I could use 
after permeabilizing the cells. However, it would be more interesting if I 
could keep the membrane structure of my cells intact, and measure in one run 
of the cell sorter (FACS) the activity/presence of CAT and the activity of the 
gene product of interest.

	Is anyone familiar with immunodetection of CAT using FACS analysis, or do 
some of you know about fluorescent substrates for CAT detection.

	Please post to this newsgroup, or mail me at the address given below. I'll 
summarize it all, promised!

	Thanx,

	Nick Vercammen
	Laboratory of Molecular Biology
	University of Gent
	Belgium
	E-mail: Nick at lmb1.rug.ac.be



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