Hi flowers: I would appreciate it if you could give me some advice on the following sort/analysis problem. Analysis will be done on a FACscan with one laser. We are staining human tumor nuclei isolated from paraffin blocks (a la Hedley): 1. primary unlabelled rabbit anti-human p53 secondary FITC-conjugated swine anti-rabbit 2. propidium iodide (PI; for nuclear DNA content) Combination 1 plus 2 has been successful. Now we also want to simultaneously measure BCL6. 3. mouse monoclonal anti-human BCL6, unlabelled (anti-BCL6 is not commercially biotinylated or labelled, to my knowledge) What fluorochrome on what polyclonal species antisera do you recommend? Or alternatively, has anyone actually used FITC, PE, 7AAD to detect 2 protein molecules and DNA content/ploidy? Is 7AAD inferior to PI for nuclear DNA content/ploidy? Thanks. Howard Howard Ratech, M.D. Albert Einstein College of Medicine/ Montefiore Medical Center 111 E 210th St Bronx, NY 10467
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