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

[Delatour Benoît]

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



   Hi,
       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
 Alonso


   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!
   B.Delatour

References

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