Gel Shift Assay Quantitation?

Ambal MD ambal at sprynet.com
Wed Sep 10 17:17:42 EST 1997


Hi Everyone!
I'm doing gel shift analysis (electrophoretic mobility shift assay) to
detect transcription factors.  I'm getting pretty good gels with
specific bands (that is, bands which disappear with excess of an
unlabelled probe similar to the labelled one, and bands which do not
change with excess of non-specific unlabelled probe).  My bands are
different in density from one nuclear extract sample to another (on the
same gel, sample loading similar). Can this be used to quantitate the
amount of the transcription factor in each sample by densitometry? If
not, how can I estimate differences in signal transduction at the level
of the transcription factor, that is, just before interaction with the
nuclear DNA?
Thanks for your reply!
--
Ambal MD
UAB
Birmingham, AL
http://home.sprynet.com/sprynet/ambal
Email : ambal at uab.edu





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