Request info. on Gel Bandshift (Retardation ) Assay
khan at ee.ualberta.ca
Thu Jul 15 00:22:18 EST 1993
I am borrowing this person's ID to post my problem.
I am a summer student doing research in the department of nephrology/
immunology. I am currently having difficulties running a bandshift assay.
Instead of seeing sharp and distinct bands of protein and DNA complex,
I am getting a smear.
We are running an 80:1 acrylamide to bisacrylamide, 5% gel.
The protein we are using is from a nuclear extraction of Jurkat and
peripheral blood lymphocytes. The method of nuclear extraction is followed
according to Dignam's protocol. Our probe was originally synthesized on
a DNA synthesis machine and purified via a trityl column. We are attempting
to purify our probe via cloning and/or an agarose gel. I would like
suggestions on improving this experiemental method.
If anyone can help me (or direct me to a more appropriate user group),
please post a follow up article or email me directly via this ID.
khan at eigen.ee.ualberta.ca
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