Dear flowers, I'm currently trying to detect surface antigens (CD4, CD8, CD45R0, CCR7 etc.) together with intracellular Ki67. For this purpose, I fixed and permeabilised cells with 1% PFA + 1% tween 20 for 15 minutes at 37°C (as indicated by Lempicki at al., PNAS, 2000) but the problem is that the fluorescence of the surface antigens is reduced a lot after fix and perm. I used also some permeabilisation kits on commerce but the protocol above gave the best result for Ki67 detection. Do you know any tricks to avoid fluorescence loss (like diminish the concentration of tween 20, stain for surface after fixation and permeabilisation....). Thank you very much, Enrico Enrico Lugli Flow Cytometry Unit Immunology Section Dept. of Biomedical Sciences University of Modena and Reggio Emilia 41011, Modena, Italy Tel. +39 059 2055425 e-mail: lugli.enrico@unimore.it, elugli@hotmail.com _________________________________________________________________ Bolletta del telefono pesante? Risparmia con il nuovo Messenger http://imagine-msn.com/messenger/launch80/?locale=it-itReceived on Tue Aug 8 11:18:00 2006
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