|From:||Gayle Callis <firstname.lastname@example.org> (by way of Histonet)|
We have not noticed any differences with the CD markers we use on murine
tissue or with huge bovine lung, unperfused with OCT or cryoprotected, and
had excellent results with cell staining.
You did not say what markers you are detecting? also how you fix, etc?
Latter could be major factors. We air dry tape transferred frozens, same as
we do with regular frozens, fix in acetone after 30 min - overnight drying,
or with acetone/alcohol mixture for markers that withstand this fixation,
or a fixative of choice.
We never go DIRECTLY into fixative INSIDE the cryostat for our murine CD
marker work or hold slides in cryostat BEFORE fixation, air drying at RT is
critical after section tape transferred to polymer slide. This is contrary
to mfr instructions, but not to our murine frozen IHC protocols already in
place. Different strokes, folk!
Am very careful to make sure antibody covers section, using a pap pen, as
the polymer is a bit resistant to good sheeting action with antibody aliquot.
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
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