liquid Bgal assay
dave eide
deide at d.umn.edu
Fri Mar 24 08:22:47 EST 1995
Hi Kathy,
Tropix sells a luminescent B-gal substrate called Galacto-Light. I've
not used it but their literature claims that it is 1000-10000x more
sensitive than ONPG. Sounds like it might be of use to you. Tropix is
at 1-800-542-2369.
Dave Eide
P.S. Check also BioTechniques 14: 762 (1993)
Hello Kathy,
I also have been wondering how to do liquid beta-gal assays on low activity
hybrids, and I was about to post a request on the yeast Net when I saw your
posting. I have some interactions in my lab that are barely detectable; some
days they are there and some days not. One suggestion that was made to me
was to get a reporter strain with multiple tandem copies of the Gal4 binding
site (a specific 17-mer) upstream of the lacZ reporter gene. The standard
two-
hybrid strains (HF7c from Clontech and Y153 form Elledge's lab) have just one
copy of the binding site. Increasing the number to 4 increases the chances
that the hybrid will bind to the promotor and activate the reporter.
***(I thought Y153 had 4 binding sites)***
I do not know anybody who has such a strain, but I suspect that it is out
there. I was going to post a request for such a strain, but I think I will
wait to see what your posting brings up. If such a strain is found, or if
some other solution to the problem of weak interactions is offered, could you
please EMail me directly? as well, you should post a summary to BioNet to
satisfy Francis.
More information about the Yeast
mailing list