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Reagent A
60ul Formic acid
50 ml H2O
Reagent B
15.6mM Sodium Carbonate (anhydrous)
68.2mM Sodium Chloride
60.5mM Sodium Sulphate
(In H2O)
Reagent C
1% Paraformaldehyde in Phospate Buffered Saline
(although this could be in just about anything, including water)
Method:
To 50ul whole blood (I've tried this with EDTA,Citrate,Heparin), add:
300ul Reagent A - vortex for 10 Seconds,
then:
132.5ul Reagent b - vortex as above
then:
50ul Reagent C - Vortex as above
Samples may be stored on ice until analysis (note-no wash step)
I've used this technique on 10s of thousands of samples and
i) it gives v.good FS/SS plots (lymphs,monos,neuts can be easily seen)
ii) it works for the following:
CD11B/CD62/CD10/CD4/CD8/CD14/CD16 oh, and lots of others I can't
remember off the top of my head.
It also works with the DCF assay.
All the best
Arnold
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web