fixation method to preserve adhesion molecules for immunostaining?

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From:Leslie Ratkay <lratkay@qlt-pdt.com>
To:histonet@pathology.swmed.edu
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Dear Histonetters,
 Could you please help me out with some fixation questions?
I need a DETAILED recipe for making up a periodate-lysine fixative and if
anybody out there using it I would also appreciate some suggestions for how to
use it (i.e. length of fixation). I will be fixing mouse ear for
immunohistochemistry. I am trying to stain for adhesion molecules CD44, CD54,
CD49d, CD106, CD62-L in inflamed mouse tissue. I can try out four different
fixation methods. I thought I could be comparing cryosectioning with/without
post fixation and paraffin embedded tissue after Zinc-Tris,
Periodate-lysine-paraformaldehyde and B5 fixation. Zinc-Tris proved to be very
effective in CD4, CD8a, CD3 and macrophage staining, however I did not have a
lot of success with the above adhesion molecules.
I would appreciate any input or suggestion of fixatives for adhesion molecules
and I will post the results in exchange.
Thank you for your help:

Leslie Ratkay
Research Scientist
QLT PhotoTherapeutics Inc





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