mouse on mouse antibodies

From:Gayle Callis (by way of histonet)

A gray area at times.

Have used the Scytek kit (check out website) for frozen sections, but did
not use their peroxidase blocker, it ate frozens off the slide.
Substituted a Dako peroxidase blocker for frozens was much friendlier.  The
kicker with this kit, their special blocker is excellent, and you have to
use their chromogen with this kit.
What really worked for us was to use their two blockers (not the peroxidase
block though),a biotinylated primary in place of a secondary antibody with
the Scytek kit since we did not have a purified primary on hand. We also
diluted out the biotinylated primary further, so do a more extended
dilution panel, this reduced our cross reaction background.

This is the basis of the DAKO ARK kit, biotinylating the primary, some
antibodies don' biotinylate as well, a potential problem, but may work with
yours, so another option - DAKO ARK kit.
There are others on the market, Zymed, Innovex, Vector MOM kit, etc, all
worth a try. Have heard some mixed reviews on Mouse to mouse kits, that
some kits work better with some antibodies, that kind of thing.  So it is a
bit of potluck.

I would use the Scytek again, it was very user friendly.

Gayle Callis
Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
Bozeman MT 

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