Hi there: We have been doing indirect immunofluorescent surface staining for phenotyping (against surface markers) on PBMC isolated from blood. We isolate PBMC from heparin added blood using Ficoll-hypaque (Pharmacia) gradient centrifugation method. Now our lab is considering using whole blood staining method since lot of others suggested that we might be losing some minor cell sub-populations during Ficoll-centrifugation and recommended whole blood staining. Since I have not done whole blood staining method so far, could people who are already doing whole blood staining method tell me where I can get the protocol (any useful references), any technical tips in adopting the method, any tips on making the lysis reagent (buffer). I would appreciate very much any help on this methodology. Thanks in advance Sathi ------------------------------------- e-mail: sathy@oitunix.oit.umass.edu
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