[Histonet] RE: Antigen retrieval question

[JR R]

Perhaps the antigen is soluble and diffused out of the
tissue into the buffer?

 

Jerry Ricks

Research Scientist

University
 of Washington

Department of Pathology

 

histonet <@t> lists.utsouthwestern.edu

 

 

Perhaps reversal is not the right word...slides were left overnight in

buffer w/ tween. We got very minimal faint staining compared to previous

runs done in one day.

 

Brett 

 

-----Original Message-----

From: Morken, Tim [mailto:Timothy.Morken <@t> ucsfmedctr.org] 

Sent: Tuesday, October 20, 2009 2:29 PM

To: Connolly, Brett M; histonet <@t> lists.utsouthwestern.edu

Subject: RE: Antigen retrieval question

 

Brett, how long? I've left them overnight before proceeding without any

problems. 

 

It would not be "re-fixation" as with formalin. However, if it is in a

detergent buffer it might cause some other kind of denaturation.

 

 

Tim Morken

Supervisor, Histology / IPOX

UCSF Medical Center

San Francisco, CA  

 

 

-----Original Message-----

From: histonet-bounces <@t> lists.utsouthwestern.edu

[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of

Connolly, Brett M

Sent: Tuesday, October 20, 2009 11:22 AM

To: histonet <@t> lists.utsouthwestern.edu

Subject: [Histonet] Antigen retrieval question

 

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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