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Hello again,
I'm in need for information about crosslinking Fc gamma receptors on cells
(human monocytes) and analyzing for H2O2 and O2 production using
DCFH-DA (Diclhorofluorescein Diacetate) and HE (hydroethidine) oxidation
assays. Any information on the reaction volume, stimulation time, etc...
or any part of the assays, would help. We get great response to PMA, but have
very unstable results with our crosslinker. We have a 30 min stimulation
with the crosslinker, and we were wondering if the oxidative production
might be on the downside of detection (would this be a correct assumption?)
Should we cut the reaction time down to catch more fluorescence from the
oxidizing fluorochormes?. Any input on this would save us some headaches.
Thank you in advance.
Sincerely,
Davin Jutila
DAVIN JUTILA, DIRECTOR SIU SCHOOL OF MEDICINE
FLOW CYTOMETRY FACILITY-1220 SPRINGFIELD, IL 62702
PHONE: (217)-524-5812 FAX: (217)-524-3227
EMAIL: SM0973%SPRINGB@SIUCVMB.SIU.EDU
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