[Histonet] Re: Hydrogen Peroxide quenching

[Edwards, R.E.]

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 <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Ford, Judi
Sent: 26 September 2007 23:04
To: Rene J Buesa; Daivik_Shah <@t> rush.edu; histonet <@t> 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 <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, September 26, 2007 2:49 PM
To: Daivik_Shah <@t> rush.edu; histonet <@t> 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 <@t> 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
_____________________________
Daivik Shah, Histotechnologist, M.S. Biotechnology RADC Laboratory, Cohn Building 436 Rush University Medical Center Tel. 312-563-4891


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