We have published a method recently using PKH-26 to label targets and
assess cytotoxicity with TO-PRO-3 (this requires a red source for
excitation). The advantage is that there is no spectral overlap. The
relevant references are:
Lee-MacAry et al: Development of a novel flow cytometric cell-mediated
cytotoxicity assay using the fluorophores PKH-26 and TO-PRO-3 iodide. J
Imm Methods (2001) 252, 83-92
Wilkinson et al. Antibody-dependent cell-mediated cytotoxicity: a flow
cytometry-based assay using fluorophores. J Immunol Methods (2001) 258,
183-191
Derek
> Our lab would like to use flow cytometry for the measuring natural
> killer cell cytotoxicity. Does anyone have a working protocol that can
> share with us? Also, could we use 24 hrs. old human whole blood
> instead of PBMC for the assay? It will be really appreciated if you
> can provide us with valuable pointers. Thanks in advance.
Lee Lam
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Derek Davies Voice: (44) 020 7269 3394
FACS Laboratory, FAX: (44) 020 7269 3100
Cancer Research UK, e_mail:derek.davies@cancer.org.uk
London Research Institute, mobile: 07790 604112
44 Lincolns Inn Fields,
London, UK.
Web Page: http://sci.cancerresearchuk.org/axp/facs/davies/index.html
In tenebris lux
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