UV

[Karen Kroeger]

I am currently trying to determine the size of a DNA-binding protein using
UV-crosslinking.  I bind by nuclear extract to a BrdU incorporated 32P
oligo, run reaction on a EMSA, then UV irradiate the gel, cut out the
complex, elute then run the elution on a SDS gel.  The problem is I get
fuzzy bands, often many bands in one lane running as a streak or smear.
The complex runs low on EMSA, but gives several high MW proteins on SDS,
which runs on SDS with a smear. How can I get clearer bands on SDS.  Any
help would be appreciated.
Thanks.  Reply to email address please.

Karen Kroeger
Dept. Biochemistry
University of Western Australia
W.A., Australia
Email : karenmk at cyllene.uwa.edu.au