Hi Phebe The first place to look when working up Antibodies is the Specification sheet from the manufacturer. If you look at the BD sheet for their CD31 Immunohistochemistry is recommended for Zinc fixed paraffin sections and acetone fixed frozeb sections but not recommended for formalin fixed paraffin sections. regards Tony
Tony Reilly Chief Scientist, Anatomical Pathology Pathology Queensland-PA Laboratory _________________________________________________ Clinical and Statewide Services Division| QueenslandHealth Level 1, Building 15,Princess Alexandra Hospital Ipswich Road,WOOLLOONGABBA Qld4102 Ph: 07 3176 2412 Mob: 0402 139411 Fax: 07 3176 2930 Email: tony_rei...@health.qld.gov.au Web: www.health.qld.gov.au/qhcss/ >>> "Phebe Verbrugghe" <pverbrug...@meddent.uwa.edu.au> 23/02/2010 2:09 pm >>> Hi Adam, Thanks a lot, might just give it a go on just formalin fixed tissue first. Phebe ________________________________ From: Adam . [mailto:anonwu...@gmail.com] Sent: Tuesday, 23 February 2010 12:01 PM To: Phebe Verbrugghe Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Double labeling with antibodies that need different fixatives I have no idea if it would work with regular formalin. In my experience, many antigens work as well in zinc buffered formalin without any antigen retrieval as regular formalin with antigen retrieval. But really, you just have to try it yourself. On antibodies I've gotten this to work, I use commercially available zinc buffered formalin which comes in gallon jugs for around $50. We don't do any special processing. We plop our sample (bones) in zinc formalin overnight at 4C, decalcify in EDTA or formic acid (only necessary for bones), and then embed just like any other sample. Adam On Mon, Feb 22, 2010 at 9:45 PM, Phebe Verbrugghe <pverbrug...@meddent.uwa.edu.au> wrote: Hello Adam, Thank you very much for this very useful information! Do you know whether this would also work on tissue fixed with formalin instead of zinc buffered formalin by any chance? Also, could you give me the recipe for the zinc formalin and can I use a standard tissue processor for embedding in paraffin or should I use a specific protocol manually and if so, which? Thanks! Phebe ________________________________ From: Adam . [mailto:anonwu...@gmail.com] Sent: Tuesday, 23 February 2010 10:52 AM To: Phebe Verbrugghe Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Double labeling with antibodies that need different fixatives Although I haven't tried it myself, others have gotten CD31 from BD to work on FFPE tissue using the tyramide amplification system on zinc buffered formalin fixed sections. I've generally had good luck with zinc buffered formalin myself for many antigens so it may work for your other one. See http://www.ncbi.nlm.nih.gov/pubmed/19052548 Just to be clear, they used zinc buffered formalin, which isn't the same thing as zinc fixative. Adam On Mon, Feb 22, 2010 at 8:41 PM, Phebe Verbrugghe <pverbrug...@meddent.uwa.edu.au> wrote: Hi all, I would like to do an immunofluorescent double labeling with two antibodies but 1 antibody works on acetone fixed frozen tissue but not on formalin fixed paraffin embedded tissue (CD31 BD pharmingen 553370) and the other one works on formalin fixed paraffin embedded tissue but not on acetone fixed frozen tissue. Is there any way I could still do a double labeling and how? Also, does anyone have experience with zinc fixative? If my antibody works on formalin fixed tissue is it likely to also work on zinc fixed tissue? 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