Dave, Calman, thanks for your fast answers. We tried to measure the beta-adrenoceptors on SF9 cells with an FACScan and a Coulter Elite ESP. One trial was to use Bodipy-CGP. We also get very nice figures with the microscope, but no chance by flow cytometry. The second trial were self made polyclonal antibodies from rabbits and the third trial were monoclonals. All the results were the same. At the moment, I have not compared the autofluorescence of SF9 cells with others. The filter settings were usual: 488 nm excitation, a 488nm blocker in the fluorescence line, a 550 nm dichroic and a 525nm bandpass for the FITC labeled second antibodies (tested for flow with other cells) and a 630 nm bp for the PI fluoresescence. I did not make the experiments with the FACScan, but they used FL1 for FITC and FL3 for PI, also usual settings. Perhaps, these SF-9 cells cannot be used for flow cytometry. Does therefore anybody knows a cell line with beta-receptors suitable for flow-cytometry? Thanks a lot Andreas ------------------------------------------ Andreas Simm Physiologische Chemie II Biozentrum, Am Hubland D-97074 Wuerzburg FAX: +49-931-888-4113 Tel: +49-931-888-4128 E-Mail: Simm@biozentrum.uni-wuerzburg.de ----------------------------------------
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