It is not a dumb question at all, every lab does things their way for a variety of reasons, the main one being of course "because we have always done it this way, and it works for us", with the real reasons why things are done in a particular way are lost in the mists of times long long ago, when the world was young and global warming had not been thought about.
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ford, Judi
Sent: 26 September 2007 23:04
To: Rene J Buesa; Daivik_Shah@rush.edu; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Re: Hydrogen Peroxide quenching
Okay, so I'm confused. Some labs do the hydrogen peroxide step before HIER and others afterwards, why? Also, are there differences using hydrogen peroxide diluted in water/methanol/or PBS? Some bring paraffin slides down to absolute alcohol then do the hydrogen peroxide step where others wait until bringing the slides to water before doing the step. Is there some rule of thumb, or do you just figure out which procedure works for that specific antibody in your lab?
I'm still pretty new to this, so sorry if this seems like a dumb question.
Judi F.
Palo Alto, CA
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, September 26, 2007 2:49 PM
To: Daivik_Shah@rush.edu; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Re: Hydrogen Peroxide quenching
You use it immediately after the HIER, before starting the IHC protocol.
René J.
Daivik_Shah@rush.edu wrote:
Hello friends,
I have question regarding Hydrogen peroxide quenching. Is this treatment applied in Immunohistostain after deparafinization or before application of secondary antibody? Is that affect any way to stain?
Thanks.
Daivik
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Daivik Shah, Histotechnologist, M.S. Biotechnology RADC Laboratory, Cohn Building 436 Rush University Medical Center Tel. 312-563-4891
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