>Hi, I have a question. The question is: What do people do to prevent >peripheral blood neutrophils from degranulating during handling and >storage. Are there special reagents, media, or buffers that can be used? >Thank you in advance, > >Victoria Boryca Smith >UNMC >vbsmith@unmc.edu Hello Victoria, Try Dulbeco's PBS +Ca/Mg 5mM Glucose *NO FCS* cells at~ 1exp6/ml and keep the cells at Room Temp (NOT on ice) I've used the above conditions to maintain PMNs for ~6 hours and the viability is good. What is more important is, I believe the method by which you obtained the cells in the first place. Prolonged contact with Dextran/Ficoll tends to get the cells reved up -I have my own method for isolation which minimises contact times -if you contact me directly I'll pass it on. Of course if you are not bothered about cell viability you can just throw truckloads of DIFP/DMEM at them 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 (0)20-7679-6234 Fax: +44 (0)20-7679-6222 email: a.pizzey@ucl.ac.uk _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/
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