Hi kevin, CPT stands for Cell Preparation Tube Apparently BD don't want to sell them to everyone, so the hide it in the maze of their complicated website. You can find them at http://catalog.bd.com/scripts/OBDsheet.exe?FNC=productlist__Alistproducts_html___362753 Only Na- Heparin and citrate are available. Price is about $5.- (per tube, not per case...) you can fin them at http://catalog.bd.com if you type CPT into the search line. Basically, it's a Vacutainer with a Ficoll soln. covered with a disgusting slimy sludge. You use it like a normal Vacutainer, then fuge it at 1500xg/RT for 20mins. That's it. You get a fine PBMC buffy on top of the sludge. You can even transport the whole thing after centrifugation with the PBMC's suspended in their own plasma. But if you plan to collect the granulocytes from the pellet via dextran sedimentation or thelike, forget it. You'll never get thru the sludge barrier. That stuff is really sticky :-( For the lysis protocol you can use Heparin- Vacutainers just as well. In Fact, I had to, since BD needed 8(!) weeks to deliver the EDTA- Vacutainers. That's why I said they'd need some competition. Carefully stick to the ratio lysing soln. / blood Good luck, brn On Fri, 19 May 2000 16:43:47 -0500, Kevin Waddick wrote: What is a Vacutainer CPT? How does it substitute for Ficoll separation? The protocol that was referred to mentioned K3 EDTA Vacutainer blood collection tubes. Are these the Vacutainer CPT? When I was in the BD website, I found about a zillion matches for Vacutainer and when I narrowed it with CPT there were far fewer but these had a relevance of 18% at most. When I looked in my BD catalog, there was no mention of Vacutainers for sale. Very puzzling. Kevin G. Waddick, Ph.D. Parker Hughes Cancer Center Bernhard Peball wrote: > 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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