Re: [Histonet] Antigen retrieval survey
Your 2 minutes would be better spent looking in an immunohistochemistry textbook. A small but excellent one is Polak, J.M. and Van Noorden, S. (1997). Introduction to Immunocytochemistry, 2nd ed. Royal Microscopical Society Microscopy Handbooks, 37. Oxford: BIOS Scientific Publications. You will find that there is an optimal technique of antigen retrieval for each antigen that has been critically studied. Some conditions (such as pH6, close to 100C for an hour) are OK for many antigens. Some require more alkaline solutions (eg pH9, more section losses!) and a few respond best to heating in a more acid (eg pH2) solution. With lower temperatures (eg 80C) longer times are generally needed. All sorts of chemicals have been included in antigen retrieval solutions, often without obvious reasons or explanations. There are published papers that compare retrieval conditions for antigens of importance in diagnostic pathology. Retrieval can sometimes be achieved without heating, as with proteolytic enzymes or 3M urea. With a survey you may find out which antigen retrieval methods are used by most of those who reply, but you will not learn anything about how to choose and use the methods, or why their discovery about 25 years ago was an important technological advance. Check out this classic paper with Web of Science, Scopus, or Google Scholar: Shi, S.-R., Key, M.E. and Kalra, K.L. (1991). Antigen retrieval in formalin-fixed, paraffin-embedded tissue: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections. Journal of Histochemistry and Cytochemistry 39:741-748. The PDF can be downloaded for free. This paper has been cited by thousands of other publications. The titles of recent citing articles may help you find a good retrieval procedure for the antigen that you need to detect immunohistochemically. John Kiernan UWO, London, Canada = = = On 23/06/15, Craig volle...@gmail.com wrote: Hi, I am conducting a short 2 min survey for my science/business class examining current trends for antigen retrieval also known as heat induce epitope retrieval. Response will be greatly appreciated! https://www.surveymonkey.com/s/7989LKR Best, Craig Vollert Graduate Student Department of Pharmacological Pharmaceutical Sciences SR2 521B College of Pharmacy University of Houston ___ 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] Antigen retrieval survey
Hi, I am conducting a short 2 min survey for my science/business class examining current trends for antigen retrieval also known as heat induce epitope retrieval. Response will be greatly appreciated! https://www.surveymonkey.com/s/7989LKR Best, Craig Vollert Graduate Student Department of Pharmacological Pharmaceutical Sciences SR2 521B College of Pharmacy University of Houston ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Antigen retrieval survey
Citrate buffer René J. --- On Wed, 7/13/11, Andrea T. Hooper andreahoo...@rocketmail.com wrote: From: Andrea T. Hooper andreahoo...@rocketmail.com Subject: [Histonet] Antigen retrieval survey To: Histonet histonet@lists.utsouthwestern.edu Date: Wednesday, July 13, 2011, 11:10 PM Hi All, I am doing a survey and will be happy to compile results and share if folks will respond! What is your favorite antigen retrieval method and/or panel? Buffer Source/composition Temperature Device Thanks, Andrea ___ 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
RE: [Histonet] Antigen retrieval survey
I hate to be pessimistic, but different clones of monoclonal antibodies can have different optimum retrieval requirements, not to mention tissue type, source species, etc. What I do believe however, is that enzymatic retrieval is under-sold - no equipment purchase is necessary so you will not see companies advocating their use. Nearly half of the antibodies we use are optimally retrieved with enzymes (pronase, trypsin, pepsin) rather than heat retrieval. Tresa Goins Veterinary Diagnostic Lab South 19th and Lincoln Bozeman, MT 59718 406-994-6353 - phone 406-994-6344 - fax Tresa -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Andrea T. Hooper Sent: Wednesday, July 13, 2011 9:10 PM To: Histonet Subject: [Histonet] Antigen retrieval survey Hi All, I am doing a survey and will be happy to compile results and share if folks will respond! What is your favorite antigen retrieval method and/or panel? Buffer Source/composition Temperature Device Thanks, Andrea ___ 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
RE: [Histonet] Antigen retrieval survey
96 degree water bath for half an hour in either citrate buffer (pH 6) or Tris-HCl (pH 9), depending on which gives better retrieval for that antibody. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Andrea T. Hooper Sent: Wednesday, July 13, 2011 11:10 PM To: Histonet Subject: [Histonet] Antigen retrieval survey Hi All, I am doing a survey and will be happy to compile results and share if folks will respond! What is your favorite antigen retrieval method and/or panel? Buffer Source/composition Temperature Device Thanks, Andrea ___ 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] Antigen retrieval survey
Hi All, I am doing a survey and will be happy to compile results and share if folks will respond! What is your favorite antigen retrieval method and/or panel? Buffer Source/composition Temperature Device Thanks, Andrea ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
