UV crosslinking

Martin Offterdinger a8803349.nospam at unet.univie.ac.at
Fri Jul 10 03:18:18 EST 1998


On 10 Jul 1998 03:57:40 GMT, karenmk at cyllene.uwa.edu.au (Karen
Kroeger) wrote:

>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

Hi Karen!
In my experience it is always a problem putting DNA on a protein gel.
Smearing and undistinct protein bands very often arise. The only
solution I can imagine is getting rid of the DNA before putting the
proteins on the gel (Dnase-treatment?).
Martin Offterdinger
Internal Med.I,Dept. Oncology
University of Vienna
Austria
E-Mail:a8803349.nospam at unet.univie.ac.at
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