Hi All! I was wondering if anyone has any experience sorting the breast cancer cell line MDA-MB-231. We are trying to sort GFP positive cells and are having a real problem with the cells agglutinating and settling out of solution. These are not your usual cell clumps associated with dying cells and free DNA as we can vortex them and get them back into suspension, they do not appear to be irreversible clumps. The usual remedies of DNAse and collagenase cocktails with the trypsin for dissociation seem not to have helped at all. Neither does passing them through nylon filter. It happens on the non-transfected cells as well. They are in DMEM w/o phenol with 5% BSA. Any ideas or advice would be most welcomed. Joanne Lannigan, MS Director, Flow Cytometry Core Facility Jordan Hall, Room 7067 P.O. Box 800734 Charlottesville, VA 22908-0734 Office: 434-924-0274 Lab: 434-243-2695 Fax: 434-982-1071 email: joannelannigan@virginia.edu
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