I use Trypsin only as a last resort. I use Pepsin because it is more gentle (does not chew up the sections). I do not use Proteinase K unless I absolutely have to and for just a few minutes as it will eat the sections off of the slides. If I use Citrate Buffer I heat it up in the microwave first then quickly put the slides in it cover it and let it sit on the bench top until cool about 20 minutes. As to why we use Citrate buffer there has been several discussions about that on the histonet. You might want to search the archieves and see what others have had to say.
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 swainfranc...@uams.edu email -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Margaryan, Naira Sent: Friday, March 27, 2009 2:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Trypsin versus other AR-s Hi Dears, It is Friday, but not a weekend yet:-):-( I just got a request from my PI to figure out: 1. How often now day's people use Trypsin (EDTA, Proteinase K or E) as an Antigen Retrieval for FFPE. 2. Why or is the Citrate Buffer pH6 more usable??? 3. Is Trypsin very old technique? Any educational feedback is appreciated, Naira _______________________________________________ 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
