endogenous peroxidase blocking

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
To:histonet@pathology.swmed.edu
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What I do depends on whether or not the sections are paraffin or frozen

Glucose oxidase/glucose/sodium azide for frozen sections or DAKO peroxidase
block containing a tidge of azide and 0.03% H2O2 in buffer. Neither of
these has affected HRP substrate after good rinsing, in fact, using a more
sensitive DAB or AEC, improved staining and antibodies had to be retitered
for lesser concentrations.  

For paraffin, depends whether detecting CD markers - I usually avoid
methanol.  Hydrogen peroxide in PBS is preferable there, and have seen H2O2
concentrations range from 0.5% up to 3% with timing around 5 - 10 min.  








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



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