Using the binding buffer coming with the APOPTEST-FITC kit of NeXins Research, the washing step will
not result in a significant decrease of cellular bound Annexin V-FITC.
Doing this you will miss a possibly interesting subpopulation, namely those cells that are about to enter
the secondary necrotic subpopulation. Those cells start to lose plasma membrane integrity without
having a fully lost barrier (a free passage of PI to the intracellular DNA) and represent a late phase of
apoptosis.
So, it'll depend on your question whether or not to include a washing step following Annexin V-FITC and
propidium iodide staining.
Chris Reutelingsperger
CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web