[Histonet] Re: MEthanol-free floating (Alonso Martinez-Canabal)

Delatour Benoît benoit.delatour <@t> u-psud.fr
Tue Jan 8 12:44:01 CST 2008

From: Alonso Martinez-Canabal [[1]mailto:alonso.martinezcanabal <@t> utoronto.ca]
Sent: January-07-08 1:27 PM
To: '[2]histonet-bounces <@t> lists.utsouthwestern.edu'
Subject: MEthanol-free floating

       I normally work with free floating IHC In brain tissue. Normally I
quench peroxidase activity with H2O2 0.3% in PBS during 30min. Allegledy,
even 5min in the normal 5% concentration would cause damage because free
floating sections are a little bit brittle. But what about the use with
methanol to protect the tissue? I heard that methanol is used to prevent
tissue damage can it be used with free floating sections?
 Thank you

   Hi Alonso,
   I  used  for  years  this  protocol on free floating sections, without
   significant damage to the tissue:
   -incubate  tissue in this quenching solution for 10 min: 2 ml methanol
   +8ml H2O dist. + 300 µl of 30% H[2]O[2]
   -rinse  5 min in H20 dist and then in buffer (eg PBS) till all bubbles
   are removed.
   This treatment is very efficient to block endogeneous perox. activity
   Good luck!


   1. mailto:alonso.martinezcanabal <@t> utoronto.ca
   2. mailto:histonet-bounces <@t> lists.utsouthwestern.edu

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