dbell at qnis.net
Wed Jan 16 13:07:42 EST 2002
On Wednesday, January 16, 2002 9:06 AM, Sophie [SMTP:sweiss at icr.ac.uk] 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.
I have not done many westerns, but I have experienced the frustration of acrylamide gone bad. It can lead to smeared samples.
Old TEMED and PMS can also effect the PAG polymerization.
If possible, I would suggest that you try staining the PAG to make sure the problem is in that stage of your work.
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