Joan, There is a detailed spectrophometric method of measuring H202 formation in the Current Protocols in Immunlogy, Chapters 7.23.12. As for phagocytosis you can use flourescent microscopy after labeling the target cells with a fluorescent dye such as PKH26 etc. The tricky part is being able to differentiate between internalized and non-internalized particles. Molecular probes sells a phagocytosis assay Kit (V-6694) that is read on a fluorescent plate reader using ~480 nm excitation, ~520 nm emission. Non phagocytozed particles are quenched by Trypan Blue. John Waitumbi ------------------------------------------------------------------------ Dr. John N. Waitumbi c/o Dr Jose A. Stoute, USAMRU, Unit 64109, Box 401 APO AE 09831-4109 Kenya Phone +254 35 22942 Email:waitumbi@net2000ke.com Fax +254 35 22903 Electronic Fax Service: 1-603- 947-4913 ---------- > 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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