NK by flow

Antony Bakke bakkea at ohsu.edu
Tue Sep 19 16:54:05 EST 2000


Look up:

Papa S and Valentini M.  Functional NK assays using flow cytometry.  Methods in Cell
Biology, 42: 193-206, 1994.  This procedure uses PI to detect dying cells.

Wierda WG, Mehr DS and Kim YB.  Comparison of fluorochrome-labeled and 51-Cr-labeled
targets for natural killer cytotoxicity assay.  J Immunol. Methods, 122: 15-24, 1989.
This procedure uses bis (carboxyethyl) carboxyfluoroscein to label targets and determines
the loss of labeled cells.

I modified the second procedure using fluoroscein diacetate succinimide ester (from
Molecular Probes)  as the label, since it covalently attaches to proteins and does not
leak out of the target cells.  This was more sensitive than 51-Cr release especially
with NK resistent targets.

Tony Bakke, PhD
Oregon Health Sciences University

>>> Bob Donahoe <rdonaho at emory.edu> 09/15 12:44 PM >>>

I'm hoping that someone can direct me to a viable protocol for measuring
NK-cell activity by flowcytometry.  And, how does such measure-up to the
standard chromium release assay?  Thanks

Robert Donahoe
Laboratory of Psychoimmunology
Emory University






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