[Histonet] Antigen retrieval question

[Rene J Buesa]

Consider the following sequence:
1- the NBF corsslinked the antigens making them "more" stable, "insolubable".
2- you did HIER and that crosslinkage disappeared, making the antigen "vulnerable" or able to be dissolved
3- you left them in buffer while "vulnerable". Does not sound to you as if the antigens were "diluted", "dissolved" in the buffer causing a weaker reaction than usual?
That would be my explanation, but to your original question ("Have you experienced...") my answer is no, I always proceeded with the IHC test immediately after HIER. Maybe that is why it has not happened to me.
Just a thought!
René J.

--- On Tue, 10/20/09, Connolly, Brett M <brett_connolly <@t> merck.com> wrote:


From: Connolly, Brett M <brett_connolly <@t> merck.com>
Subject: [Histonet] Antigen retrieval question
To: histonet <@t> lists.utsouthwestern.edu
Date: Tuesday, October 20, 2009, 2:21 PM


Has anyone experienced a "reversal" of citrate HEIR unmasking do to a
prolonged delay in continuing the experiment after the retrieval step? 

Brett M. Connolly, Ph.D.
Research Fellow, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
tel. 215-652-2501 fax. 215-993-6803
brett_connolly <@t> merck.com

  
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