Brett, We mostly work & do IHC on 40um free-floating brain sections. These sections were cut from fixed brain blocks. Sometimes, not on a regular basis, but sometimes we have had to place our sections in (plain) PBS solution & stored at 4C - overnight before continuing the IHC protocol with no effect to the staining quality. Keep in mind, that this was done before the primary antibody. We have also placed our cut and unstained sections in plate wells (5ml of PBS) - again at 4C for 2 days before starting our IHC protocol. We use quite a number of different antibodies & stain using polymer.
I hope this helps. Maria Mejia Histology Manager Department of Neurosurgery UCSF Sf, CA 94103 ________________________________________ From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Connolly, Brett M [brett_conno...@merck.com] Sent: Tuesday, October 20, 2009 11:33 AM To: Morken, Tim; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Antigen retrieval question 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.mor...@ucsfmedctr.org] Sent: Tuesday, October 20, 2009 2:29 PM To: Connolly, Brett M; histonet@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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Connolly, Brett M Sent: Tuesday, October 20, 2009 11:22 AM To: histonet@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_conno...@merck.com Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu - direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu - direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
