Hello. I`m having a HUGE problem trying to intracellularly stain primary t-cells which have been retrovirally infected with GFP. My infection %`s of GFP into extracted splenocytes are only about 5% at the beginning of my experiment, but at the end of the experiment, once I have fixed and permeabilized and stained the cells for cytokines, I can detect no GFP positive cells. I know my insert is there (as my lysates on a western showed), but how do I get around losing the fluorescence of the GFP. Please help! Are there any protocols that circumvent the quenching of the GFP signal? Thank you very much. Sincerely, Raman Minhas Clinical Research Institute of Montreal, McGill University
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