mw132 at mole.bio.cam.ac.uk
Wed Jan 16 14:28:46 EST 2002
On 16 Jan 2002, Sophie wrote:
> I am having major problems carrying out western blots. I ALWAYS end up with
> a smear down the lane instead of a tight, compact band. I know that my
> protein isn't degraded and that the antibodies are specific to the protein
> and I have remade all my buffers. The only thing I can think of that might
> be causing a problem is the acrylamide. I read that acrylamide can last up
> to 9 months unopened and mine has been open for about 18 months, do you
> think this could be affeecting my gels?
> Any help and comments is much appreciated.
have you just overloaded the gel? When I do this I get a
horrible smear in my western. Is your sample a crude extract? Maybe
there is a lot of salt carry over from the medium: use less sample in the
gel. How are the standards looking? If they are OK, then the acrylamide
is probably OK too. Good luck, Mike.
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