SDS-PAGE gel resolution
A. F. Simpson
afs7 at le.acluk
Mon Aug 7 09:56:13 EST 2000
Martin Thompson wrote:
> I was wondering if anyone knows what kind of factors influence how
> 'fuzzy' bands are on a SDS-PAGE gel. Currently I am having problems
> with this and would like to know how to increase the sharpness or
> resolution of these bands.
Running the gel itself too fast (and too hot) can cause fuzzy bands.
Try reducing the current and seeing if things improve. Do you use a
prestained marker? That's the best way to keep an eye on the run.
Are the bands fuzzy on the gel or only after blotting? A friend had
trouble with fuzzy bands which was eventually traced to trying to run
the semi-dry blotter at too high a power. I don't know if the same
effect would occur on wet blotting. I have always taken a 'tortoise'
approach to blotting, so I've never encountered such a problem
Can you give a few more details? (E.g. type of sample, MW of the
protein you're looking for, size/type/percentage of gel, what staining
you used to visualise the 'fuzziness'.) The more detail you give the
more chance there is someone can give a helpful response.
> Thanks in advance,
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