Carolyn A. Taylor, Ph.D.
ckeever at post.its.mcw.edu
Fri Feb 17 18:58:25 EST 1995
Does anyone out there have any information
> >NK assay reliability or use of the suggested method?
> >Mary Bennett
In article <3i05rn$8t5 at netnews.upenn.edu>, David Peritt
<Peritt_d at a1.mscf.upenn.edu> wrote:>
> We conduct NK assays often using various targets and have never worried
> about the target variability. These cells have been in culture so long
> that all instabilities have been worked out. Whether cycle differences
> will matter has not bothered us much. We try to use cells in log phase
> growth for labeling.
I too have performed hundreds of NK, LAK and CTL assays and find little
variation in healthy (mycoplasma free) target cell cultures. Labelling is
better when the cells are in log phase as mentioned above, however, Monday
morning assays seem to work fine for us too. 51Cr less than 3 weeks old
and mycoplasm free target cells are two critical factors along with gentle
washing and addition of targets. By gentle I mean no undue agitation in
plating and wash steps not exceeding 1200 rpms in a table top centrifuge.
Of course accurate collection of supernatants, cell free, at harvest is
CA Taylor, Ph.d.
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