I would be most grateful for any advice/methods/references etc for cytotoxicity assays using flow cytometric techniques. Sarah, I have had very good luck in the past using standard TUNEL and/or Annexin V techniques to detect CTL-mediated apoptosis in target cells by flow cytometry. These techniques are especially straight-forward if you are using tumor cells as your target. For instance, TUNEL positivity is dependent on the presence of CTLs, can be blocked by EGTA (blocks perforin function), and can be detected as early as 5-10 minutes after the addition of CTLs to targets. I don't have any specific references on hand, but I'm sure you can get these standard techniques working in your system without much effort. Best regards, Andrew D. Wells, Ph.D. University of Pennsylvania Department of Medicine 728 Clinical Research Building 415 Curie Boulevard Philadelphia, PA 19104 (215) 573-1840 [office] (215) 898-1951 [laboratory] (215) 573-2880 [FAX] adwells@mail.med.upenn.edu
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