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Question (24229) | HistoReveal (15ml) (ab103720)

Hi! Thank you for your reply! I will try to detect myofibroblasts in cancer FFPE tissue using an antibody against alpha-SMA (ab7817) and a Alexa fluor-secondary antibody. To try to investigate the cell-type specific expression in this case I will use laser capture microdissection, extract RNA from the collected cells and run a qPCR on a small number of genes. I know that the RNA is likely to be degraded due to the fixation method but I will try and hope for the best. At least the FFPE tissue we have is fairly new and have been stored in 4°C. Most people in my lab use heat-induced antigen retrieval techiques for IF and IHC but they are not extracting RNA and I'm afraid that these protocols will affect the RNA quality. Therefore, when I found your HistoReveal I got a bit curious... Just tell me if there is any more information that you need!

Thank you for confirming the details. The HistoReveal ab103720 use in IHC is based on enzymatic antigen retrieval method. As there is no step of heating the tissue sections so probably this could be product as per your requirements. However I cannot be 100% sure because we have never tried this product in our own lab in similar kind of experiment so we will not be able to share any data regarding this. Please be advised that the optimization of experiment has to be done by the end user. I would also suggest searching the literature about the common method other scientist are using. I have done quick search which I am happy to share; http://jhc.sagepub.com/content/49/8/931.full.pdf+html Masuda N, Ohnishi T, Kawamoto S, Monden M, Okubo K (1999) Analysis of chemical modification of RNA from formalin-fixed samples and optimization of molecular biology applications for such samples. Nucleic Acids Res 27:4436–4443 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC148555/pdf/270e12.pdf I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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