bernard at elsie.nci.nih.gov
Thu Jun 16 00:59:25 EST 1994
In article <MAILQUEUE-101.940615165156.384 at vmp.vetmed.wsu.edu>, GCANTOR at VETMED.WSU.EDU ("Glenn Cantor") writes:
> I have transfected cells with a plasmid containing a hygromycin
> selectable marker. To compare transfection efficiency in various
> cell lines, I would like to compare the amounts of hygromycin
> phosphotransferase which the lines express. Does anyone know if
> antibodies or an antibody-based kit are available for detection of
> hygromycin phosphotransferase? (5'-3' has an ELISA for detection of
> neomycin phosphotransferase II, but doesn't have one for hygromycin.)
> Debra Hoover
Why do you need the antibody? Wouldn't it be sufficient to test the
hygromycin sensitivity of the transfected lines, or are there complicating
factors? Alternatively, how about cutting out the hyg gene from the plasmid
and using it to probe Northerns?
While we are on the topic, has anyone any experience of cells
becoming hygromycin resistant without transfection with this gene? I believe
that G418 is not a substrate for classic multidrug resistance (pgp/P170/mdr)
but I do not know if hygromycin has been tested.
Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)
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