Whilst looking through a message board in the past, I remember seeing an
archive describing a neutrophil phagocytic assay. I know that there are kits
on the market such as Phagotest(r), but I would like to have a go at an
assay myself. The plan I have is to use fluorescein conjugated zymosan
particles as the target and hence measure phagocytosis on the flow
cytometer. I read that you can quench mebrane bound and free label with
propidium iodide, however, I would prefer to have a control in which I add
cytochalasin D, which inhibits phagocytosis. Does anybody have a protocol
for a similar method? I need to know measurements such as the ratio of
neutrophils to zymosan, and the amount and time of incubation for
cytochalasin D. Do I need to attach complement to my zymosan? How is best to
do this? (time and temperature of incuabtion etc.) Also, would it be
possible to carry this assay out on whole blood (maybe with a RBC lysis
step)?
Many thanks
Daniel G. Blount
Nottingham University
School of Pharamceutical Sciences
University Park
Nottingham
NG7 2RD
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