I always found that ACK lysis buffer was gentle, and very inexpensive compared to some of the commercial preps (which also work well too). DO a search on the archives, and you will find lots of info there. Otherwise, I believe that "Handbook of Flow Cytometry Protocol's" (? is that the name?) authored by J. Paul Robinson's, and perhaps someone else, but cannot remember (and I am writing this from home)is an excellent resource. When searching, look under "lysis". If you have trouble,email me privately, I will dig into my box of disks and try to find the draft for the SOP I wrote. P. Echeagaray >From: Sathi <sathy@vasci.umass.edu> >To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> >Subject: Surface staining on whole blood samples >Date: Tue, 16 May 2000 12:19:54 -0400 > > >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 > > ________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com
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