Sorry away for the weekend, so missed the start of this thread. All the
ideas offered so far are good methods for measuring hydrogen peroxide.
Phagocytosis per se can only measured by incorportation of labelled target
cells by effector cells. The best method is debatable. Radioactive labels
are better because there is less temptation to count a field of view that
looks good. On the otherhand its radioactive, which is a real downer on use
and disposal.
You can use superoxide and other free radicals (especially H2O2) as a
measure of phagocytosis in PMN, because this has been shown to be dependent
on internalisation and release of myleoperoxidase stores. Not sure about the
method in "Methods in Immunology" quoted by John I haven't seen the book,
but if a luminometer is available then using luminol is a good method. You
can also use cytochrome c to measure superoxide in a spectrophotometer. Both
methods have been published repeatedly, and can be found in "Methods in
Enzymology". Sorry don't have the reference to hand but the authors are
Hancock and Jones.
hope this helps.
Craig Turner
IBGRL
----------
> From: joan Kalnitsky <jkalnits@vt.edu>
> To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
> Subject: H202, Phagocytosis
> Date: Thursday, December 16, 1999 12:16 PM
>
>
> This is not a Flow question, but I thought this group might be able to
help
> out. Does anyone know of any protocols to measure H202 and phagocytosis
in
> a manner other than flow cytometry. We have a PI who has measured both
of
> these via flow and is now working in Chile where they have no flow and
> would like to be able to measure these.
> Any and all input will be greatly appreciated.
> Thanks in advance,
>
>
>
> Flow Cytometry Lab Supervisor
> VMRCVM
> (540) 231-4115
> FAX 540-231-7367
> jkalnits@vt.edu
>
> "It is better to serve than to receive."
> B. Borg
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