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I've been asked by one of my workmates about the use of screening
patient sera for anti neutrophil cytoplasmic antibodies (ANCA), by
flow. The idea is to screen out all the negatives and retest the
positives either by ELISA or IF on neutrophill slides. I know that
this is possible but I have a few queries;
a) What is the best bethod to obtain and store neutrophils for
screening (there needs to be minimal activation of the neutrophils!).
b) What is the best method for permeabilising the cells, in house or
commercial products?
and lastly
c) Best detection method: Fab2 FITC anti IGG or would it be better to
use a FAB2 biotin, followed by streptavidin FITC,
I would appreciate any ideas on these or any other aspects of this
test.
Very many Thanks
John Culkin
Dept of Immunology
Royal Liverpool Hospital
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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