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)