dbell dbell at
Wed Jan 16 13:07:42 EST 2002

On Wednesday, January 16, 2002 9:06 AM, Sophie [SMTP:sweiss at] wrote:
: Hello,
: 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.
: Thanks,
: Sophie.

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.

Deanne Bell
USDA Agricultural Research Service
9611 S. Riverbend Avenue
Parlier, CA 93648
(559) 596-2806
(559) 596-2791 fax
dbell at


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