PROPIDIUM IODIDE CELL CYCLE ASSAY

Sylvain GIMMIG cytometry at gimmig.org
Wed Jun 20 08:56:56 EDT 2007


Dear colleagues,

We have two questions:

1. We have been following a standard protocol for the PI cell cycle assay
 following stimulation
of human CD4 cells or Jurkat cells with CD3:CD28 antibody-coated Dynal beads
(Pan mouse IgG Cellcept beads). However, the cells seems to be very fragile
following fixation in 70% ethanol (either at 4C or -20C and for periods varying
from  either 1 hr to days of fixation). We are losing many due to lysis.  Does
someone have suggestions on how to reduce cell loss due to lysis at this step?

2. What are the best time points to measure in analyzing the cell cycle in human
CD4 and CD8 cells following stimulation with CD3:CD28 antibodies?

Thanks for any input,


-- 
Sylvain GIMMIG
Flow Facility Manager
HÔPITAL ST-LUC/CHUM
Bureau:409
264, Boul René-Lévesque est
Montréal, QC, H2X 1P1, Canada
Téléphone: 514-890-8000 ext 35778
FAX: 514-412-7314
Website:http://www.chumtl.qc.ca



More information about the Cytometry mailing list