Dear Sathi, I use the lysis protocol mentioned on the "BD Monoclonal Antibodies Source Book" CD. It's very simple and the viability of the cells is good. I avoid Ficoll, if possible. Too tedious. A Vacutainer CPT does the same and is a lot easier in handling. If you don't have the BD CD, you can find the protocol at http://www.bdfacs.com/source_book/html/23_1372n.shtml Anyway, if someone has a better one, I'm always open to improvements. BTW: is there any alternative to BD CPT- Vacutainers? The product is just fine, but our local BD representative could need a little competition :-) Yours, brn On Tue, 16 May 2000 12:19:54 -0400, Sathi wrote: > >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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