G0/G1 and G2/M identification
darzynk@nymc.edu
Tue, 21 Mar 1995 12:13:41 -0500
For the past 15 years we routinely use cell staining with AO (after
partial DNA denaturation) to discriminate between M and G2 or G0 and
G1 (the method is best described in Vol 41 of Methods in Cell
Biology). This method is(was) also used in many other labs. Using
FACScan which has short sample flow tubing, a 10 min wash with 10 %
bleach and with detergent (0.5% Triton X-100) is adequate to remove
any AO from the tubing. We use this instrument interchangebly
measuring immunofluorescence and AO, with no problems at all.
Mitotic cells can also be discriminated from G2 cells by the lack of
cyclin A (antibody to cyclin A is available from PharMingen).
Zbigniew Darzynkiewicz
CD ROM Vol 2 was produced by 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:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
