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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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