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band-shift assays

Stephen Nurrish s_nurrish at icrf.icnet.uk
Thu Jul 28 13:23:49 EST 1994


In article <dhs3-250794105341 at 132.236.171.142>,
dhs3 at postoffice.cornell.mail.edu (daniel schott) wrote:

> Does anyone have expertise in band-shift (protein-Dna binding) assays?  A
> friend of mine is having horrible trouble repeating a series of
> experiments,  her samples get stuck in the wells.  Due to the nature of the
> experiment, she has to use very low concentrations of DNA and no
> non-specific DNA competitor.  She doesn't think the problem is aggregation.
>  Is the purity of the acrylimide absolutely vital for this stuff to work? 
> Reply to bd14 at cornell.edu, and I'll pass on suggestions.  Thanks!  

a definative answer to this depends a lot on the source of the protein. We
use bandshift a lot for retic lysates and cell extracts, in these cases the
absence of non-specific competitor gives exactly the result you describe,
everything stuck in the wells. The usual purpose of bandshift is to show a
specific interaction between DNA and protein so why can't she use
non-specific competitor?



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