glucose oxidase peroxidase blocking

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From:Gayle Callis <>
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For anyone who is using glucose oxidase endog peroxidase quenching, would
like to know what they are getting for morphology on acetone fixed frozen
sections, either with a single acetone fixation or a double acetone
fixation with air drying step in between 1 and 2 acetone baths, 4C.  

If their sections have (in the past) and do look like my sections, ugh! a 4
x 4 truck is running over them! Chemicals were weighed out on an analytical
balance, use a 37C waterbath for 1 hour, gentle washing steps, careful
rinse after the GLUOX 3 X 5 min rinses in pure PBS, some Tween 20 in rinse
buffer, 0.05%, plus charge slides, long drying before fixation, freshly cut
sections stained the next day after careful -80C storage, no full moon,   

An occasional section looks fairly good, but the rest are motheaten!
Longer incubations (Tween used here) were worse than a short protocol (non
Tween) and double fix was better than single fix.  

A whole day in the garbage, but then what's new in the IHC world!

Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

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