Re: H202, Phagocytosis

From: John Waitumbi (waitumbi@net2000ke.com)
Date: Sat Dec 18 1999 - 16:37:44 EST


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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