[Histonet] Immuno. confusion

[Rene J Buesa]

If you fix with formaldehyde, HIER will eliminate the cross link and that step is necessary BEFORE you can apply the peroxide. The enzyme (peroxidase) is also a protein and has to be "open" to receive the H202.
The same if you want to apply enzymatic digestion, HIER has to be done first.
HIER will remove the formaldehyde cross linking and this step is required before any other in the IHC protocol.
René J.

--- On Wed, 8/3/11, Hannen, Valerie <Valerie.Hannen <@t> parrishmed.com> wrote:


From: Hannen, Valerie <Valerie.Hannen <@t> parrishmed.com>
Subject: [Histonet] Immuno. confusion
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
Date: Wednesday, August 3, 2011, 11:57 AM


Hi folks...

I am going to try to revamp out Immuno. procedure and am looking at a couple of spec. sheets from our primary vendor. I am gettin conflicting information...so I am turning to you all for help.

When doing immuno's that require either HIER or enzyme digestion, do you perform the retrieval first or do you do the "peroxide" step first??
One spec. sheets says retrieve then block endogenous peroxidase...the other says block endogenous peroxidase then do the HIER or enzyme digestion..which is correct??

One other question that I have is...what do you use to enhance your stain?? The enhancing solution that we were using has been discontinued.

Thanks so much!!

Valerie Hannen,MLT(ASCP), HTL,SU(FL)
Histotechnologist
Parrish Medical Center
951 N. Washington Avenue
Titusville, Florida 32796
(321) 268-6111 ext. 7506



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