IUBio

5 kDa protein detection

Biology jsullivan at moon.sba.dal.ca
Wed Jan 6 15:03:58 EST 1999


It is strange but try Ethidium bromide for staining the SDS PAGE, about 6
minutes is enough.

Mahmood

Ryan Werstuik wrote:

> I would try using protein markers (with your smallest at 5kDa), running
> the sample in a center lane, and 1/2 to 2/3 the current time you are using
> now.  I have also heard/read of using a stain that allows the samples to
> be seen as the gel runs; however, I can't remember the where.
>
> I hope this helps,
> Ryan
>
> Rasmus Wendelbo Nielsen wrote:
>
> > I am currently trying to purify a protein of 5 kDa, but are having
> > problems detecting the protein. As it only contains tree tyrosines and
> > no tryptophans the A280 is very low. I have tried running 18% SDS-PA
> > gels and a 15 to 25% SDS PA gradient-gels, without luck, the protein
> > probably diffuses out of the gel during staining.
> > So I am looking for other (easy) ways to detect low MW proteins. Any
> > ideas would be appreciated!
> > I have heard that adding coomassie to the running buffer instead of
> > post-staining the gels could be a solution. Anyone got experience with
> > that?
> > Thanks, Rasmus






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