Re: [Histonet] Frozens and antigen retrieval
You should not have to use retreival. Retrieval either by heat/solution or enzyme digestion is usually needed when the antigen/antibody site has been blocked/masked by the fixation process. Kim D From: Daniela Bodemer daniela.bode...@mcri.edu.au To: histonet@lists.utsouthwestern.edu Sent: Wednesday, July 4, 2012 7:02 AM Subject: [Histonet] Frozens and antigen retrieval Hi all, A question from my student: antigen retrieval when using frozens for immunofluorescence -yes or no? The protocol suggested by the antibody company lists the retrieval as an option. I am used to do retrieval on paraffin sections, but not on cryo sections. Hit me with your opinions on this :-) Thanks in advance, Daniela Sent from my iPad __ This email has been scanned by the Symantec Email Security.cloud service. For more information please visit http://www.symanteccloud.com __ ___ 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
[Histonet] Frozens and antigen retrieval
Hi all, A question from my student: antigen retrieval when using frozens for immunofluorescence -yes or no? The protocol suggested by the antibody company lists the retrieval as an option. I am used to do retrieval on paraffin sections, but not on cryo sections. Hit me with your opinions on this :-) Thanks in advance, Daniela Sent from my iPad __ This email has been scanned by the Symantec Email Security.cloud service. For more information please visit http://www.symanteccloud.com __ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Frozens and antigen retrieval
Usually not. Fixation cross-links the proteins, which can mask the epitope (=antibody binding site of the antigen). So antigen retrieval breaks the fixative cross-links, exposing the epitope. If there's no fixation, there's no cross-links, so the epitope is usually exposed and available to easily bind to the antibody. Plus, there's no destruction of tissue morphology if you're not using antigen retrieval, so the quality of the section looks much nicer. That being said, there may be some antibody out there that still needs antigen retrieval on frozen section, but then the company's protocol probably wouldn't say optional. So try it the first time without antigen retrieval. Peggy A. Wenk, HTL(ASCP)SLS Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 The opinions expressed are mine, and do not reflect Beaumont. -Original Message- From: Daniela Bodemer Sent: Wednesday, July 04, 2012 7:02 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozens and antigen retrieval Hi all, A question from my student: antigen retrieval when using frozens for immunofluorescence -yes or no? The protocol suggested by the antibody company lists the retrieval as an option. I am used to do retrieval on paraffin sections, but not on cryo sections. Hit me with your opinions on this :-) Thanks in advance, Daniela Sent from my iPad __ This email has been scanned by the Symantec Email Security.cloud service. For more information please visit http://www.symanteccloud.com __ ___ 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
