RE: ANCA by flow
Edoardo Colombo (coledoco@tin.it)
Mon, 14 Jul 1997 07:27:13 +0200
John
the best way I know to recover neutrophils is to lay
0.5 ml peripheral blood onto a 1 ml ficoll 1.077 and
let motionless for 15 minutes : you will have a clear
pellet containing all WBC and the platelets (by the
way this is a quick and simple way to obtain very rich
plt specimens); this tecnique do not activate the PMN
at all.
Regarding the permeabilization I think that Triton X-100
(working concentration = 0.1 %) is the best item you
can use
Finally I think that Fab FITC IgG , as used by me to
detect anti-platelets autoantibosies , is the right way
to detect the target you need.
Best regards
Edoardo Colombo MD
Director Flow Lab
Dept. of Pathology
Valduce Hospital
COMO , Italy
coledoco@tin.it
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