Re: Cytotoxicity assays

From: Andrew D. Wells (adwells@mail.MED.UPENN.EDU)
Date: Mon Jul 24 2000 - 09:42:22 EST


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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