Re: Tunnel Assay
Wayne Green (Wayne.Green@genetics.utah.edu)
Thu, 17 Oct 1996 09:22:09 -0600
Jim:
We have done this assay with surface staining and it worked on murine
spleen cells. The Tdt/TUNEL assay is usually done with FITC as the reporter
fluor (ie. FITC-strepavidin with biotin-dUTP). We found that there was alot of
problems with autofluorescence and spill over from FL1 that made use of PE
almost impossible for detecting surface Ag's. We therefore tried turning
things around and did our surface staining with FITC and the apoptosis was
detected with SA-PerCP; this worked nicely on the FACScan. Other than that we
used the original protocol pblished by Gorczyca et al, Int. J. Oncol 1:639-648
(1992); if your investigator is using one of the commercial kits that may behave
differently.
--
Wayne Green
wgreen@genetics.utah.edu
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