I work with non-decalcified and decalcified bone. After I have tried to pressure cook it, steam it, microwave it, etc my sections look pretty bad(coming off, etc.) I have found that either using Pepsin (Zymed) for 20 minutes at 37 degrees C. in a humidifying chamber or Pepsin for 20 minutes at room temperature in a humidifying chamber or microwave the buffer to boiling, quickly remove and place slides in solution, cover and let sit for 20 minutes or until cool to touch work the best for my purposes.
Frances L. Swain HT(ASCP) A. A. S. Special Procedures Technician Department of Orthopaedic Surgery Center for Orthopaedic Research Barton Research Building 2R28 4301 West Markham Street Little Rock AR 72205 (501) 686-8739 PHONE (501) 686-8987 FAX [EMAIL PROTECTED] email -----Original Message----- From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Sebree Linda A. Sent: Friday, October 03, 2008 12:49 PM To: Patten, Nicole (NIH/NIAAA) [F]; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Antigen Retrieval Try using a laboratory pressure cooker like the Decloaking Chamber from Biocare Medical. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -----Original Message----- From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Patten, Nicole (NIH/NIAAA) [F] Sent: Friday, October 03, 2008 12:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Antigen Retrieval Hi- I am new to using IHC and I need some advice on Antigen Retrieval. I usually do it in an autoclave for 15min at 121C for FFPE human brain tissue (in Citrate or Tris Buffer), but by the end my tissue looks pretty bad and parts have even fallen off. The tissue is fixed on SuperFrost Plus Slides. Any suggestions would be really helpful. Thanks! N. Patten Post-Baccalaureate Fellow/IRTA National Institutes of Health _______________________________________________ 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 Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
