[Histonet] peroxidase blocking in whole tissues

Andrea Marion amario3 <@t> uic.edu
Wed Jan 25 14:36:27 CST 2012


Hello histonet,

I am working on a whole-mount Pecam immunostain on mouse embryo hearts.
During the peroxidase blocking step (1% H2O2 in PBS) there was visible
vigorous bubbling in the tissue which caused the heart to swell up.

I will try a lower concentration of H2O2 and also prepare the solution in
MeOH which I have read is "more gentle" on tissues with high endogenous
peroxidase activity. My questions are 1) Why is MeOH "more gentle" than
H2O2 in aqueous solutions? and 2) Is there anyone out there in histoland
who has done this experiment and can recommend a protocol?

Thank you,

Andrea Marion
Graduate Student
University of Illinois at Chicago




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