I was surprised by Martin's experience. I buy lots of Jackson
antibodies (FITC, Cy3, Cy5, HRP; goat and donkey anti-mouse and
anti-rabbit) and generally aliquot them out as 10 ul/tube and freeze
and store at -80. Once thawed, I dilute and use that day and throw
any extra away. They have been stable for years this way. Tom
>Shinohara, Mari wrote:
>
>> I am now trying to see subcellular localization of molecules. When I use
>> FITC as secondary staining, it looks nice. However, if I use Cy3, there
>> appears pretty high background (all over, not only in cells) and the whole
>> field looks orangish as well.
>
>Have you ever run the secondary antibody through a freeze-thaw cycle?
>If so, that may be the problem. One freeze-thaw cycle, in my
>experience, turns most Jackson secondaries from being superb, to being
>rather mediocre.
>
>Good luck!
>
>Martin Wessendorf
>--
>Martin Wessendorf, Ph.D. office: (612) 626 0145
>Associate Professor lab: (612) 624 2991
>Dept. Neuroscience Preferred FAX: (612) 624 8118
>University of Minnesota Dept FAX: (612) 626 5009
>Minneapolis, MN 55455 e-mail: [log in to unmask]
--
Thomas E. Phillips, Ph.D.
Associate Professor of Biological Sciences
Director, Molecular Cytology Core Facility
3 Tucker Hall
Division of Biological Sciences
University of Missouri
Columbia, MO 65211-7400
(573)-882-4712 (voice)
(573)-882-0123 (fax)
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