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Hi! I'm going to use FFPE sections for laser-capture microdissection of cells for RNA extraction and qPCR. I'm planning to visualize the cells of interest with IF and has been looking around for a suitable antigen retrieval protocol that does not harm the RNA. Would you say that HistoReveal is suitable for this application or if you have any better suggestion? Best Regards
Asked on Sep 21 2011
Thank you for contacting us. Could you specify which target you would be interested in seeing with antibodies? I don’t think the antigen retrieval method will affect the target detection in IHC-IF. However if you are interested in extracting RNA from FFPE tissue sections then the procedure will be different. Because formaldehyde affects nucleic acids by forming adducts with exocyclic nitrogen atoms in nucleotides. This immobilization of DNA and RNA is attributed to trapping of the long nucleic acid molecules in networks of associated basic protein molecules, which are cross-linked by methylene bridges. Could you also provide more information about the experiment you are planning to do? I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Abcam Scientific Support
Answered on Sep 21 2011