lacZ detection by FACS

From: Karim Vermaelen (Karim.Vermaelen@rug.ac.be)
Date: Thu Mar 22 2001 - 06:05:39 EST


Hi,

We're planning to use a green fluorescent b-galactosidase substrate kit
from Molecular probes (Imagene Green C12 lacZ Gene expression Kit) to
detect lacZ expression in single cell suspensions. In contrast to FDG,
the ImaGene does not require hypotonic shock treatment for substrate
loading.
-> Does anyone have experience with this staining protocol in
combination with surface marker staining? Also, what would you recommend
as substrate incubation time for leucocytes (the range mentioned in the
datasheet is 20 min up to several hours).
NB the cells were not transfected in vitro, but are derived from lacZ
transgenic mice.
Thanks a lot for any advice!

Karim Vermaelen



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