We have been doing some experiments looking at markers on activated mouse platelets. Now a member of our lab wants to do a time course of expression following thrombin induced activation. The plan was to remove samples at various time points following activation, fix, and then stain all sample at the same time at the end of the time course. But we have noticed that non-thrombin treated platelets appear positve for P-selectin when we fix with 1% paraformaldehyde (unfixed are negative so we are not activating during the isolation procedure). Does anyone have a fixation method they use prior to staining platelets that does not cause them to up regulate P-selectin? thanks Robert Jensen
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