DNase FOOTPRINTING Off a Gel-Retarded Complex

[Sandy Curtiss]

In article <3bde8u$l6p at bigfoot.wustl.edu> Thiruvamoor P. Ramkumar,
ramu at artsci.wustl.edu writes:
>Hi,
>	i'm looking for help on doing DNase Footprinting from a 
>DNA-Protein complex isolated from a gel-retarded complex. Primarily, my 
>concern is to elute the DNA-Protein complex out of the gel without 
>disrupting the complex. Possible buffer compositions, elution
conditions, 
>quantitation parameters etc. will all help. 
>	
>
> _
>ramu

A protocol for doing DNase footprinting directly in a mobility shift gel
is given in Kuwabara, MD and Sigman, DS (1987) Biochemistry 26:
7234-7238.  I have not used this protocol, but it was used successfully
by Wolfe et al. (1993) JBC 268: 12418-12426.  Hope this helps.

Sandy Curtiss
Monsanto Co.
swcurt at ccmail.monsanto.com