>Dear flowers, we are trying to detect intracellular IFN-g in murine spleen cells, but we have problems with positive control. We activated 8 x 10E6 spleen cells with 10 and 20 ug PHA, and with 6 ug PMA + 4 ug Ionomicin for 4 hours but results were no IFN-g. We used 0.2 ug anti IFN-g-PE from Pharmingen. >In our lab., we activated human whole blood with 10 ug PHA for 4 h and intracellular IFN-g were detected. >Any of you have experience in murine cells activation and IFN-g detection? >Thanks in advance, >Martha Greetings Martha, I suspect that you will have to incubate your cells with somthing like Brefelden A for ~4 - 12 hours to allow accumulation of IFN-g to a level where you can detect it with you ab. I have some protocols that I can find if you would like me to send them to you. Regards, Arnold _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/ Arnold Richard Pizzey Department of Haematology Royal Free and University College London Medical School 98 Chenies Mews London WC1E 6HX U.K voice: +44 020-7679-6234 Fax: +44 020-7679-6222 email: a.pizzey@ucl.ac.uk _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/
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