b-galactosidase assay after transfection

Martin Leach leach at mbcrr.harvard.edu
Thu Apr 14 10:23:04 EST 1994


In article <199404141123.UAA14023 at inetnif.niftyserve.or.jp>,
HGF02633 at niftyserve.or.jp (=?ISO-2022-JP?B?GyRAQFBETT1TPCMbKEo=?=) wrote:

> Does anybody know sesitive assay system to detect beta-galactosidase 
> activity in cell extracts after DNA transfection (lipofectin method). 
> We transfected pCH110 (beta-galactosidase expression vector) into PC12 
> cells. However, the transfection efficiency was very low. We could not 
> detect the enzyme activity with o-nitrophenol-b-D-galactopyranoside 
> (ONPG) as a substrate, though the CAT activity was detected in the same 
> expeirment. How can we overcome this problem. please teach me.

I believe there may be a fluorescent substrate for bgal. Detection of this
can be made at very low levels - i think (please correct me)


Martin
-- 

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