From: Nunes, Perla (Perla.Nunes@carolinashealthcare.org)
Date: Wed Jan 15 2003 - 11:05:46 EST
Dear all, I am fairly new at Flow. I was running an experiment using PE conjugated antibodies and 7-AAD. Cells were surface stained with PE conj. Ab, washed, then (using Sigma's protocol) cells were further stained with 10 ug/ml of 7-AAD in PBS on ice for 30 mins. After washing with PBS twice, the cells were fixed with 2% paraformaldehyde supplemented with 50 ug/ml actinomycin D. When I ran these cells on the FACS calibur, all the cells (unstained vs. control Ab vs. + Ab) were running in FL3. I also ran another set that did not get 7-AAD but were fixed in 2% paraformaldehyde, in this set there was also no positive staining. In the past we've used BD's Facs lysing solution and the Flow data has looked great. Has anyone used 7-AAD with BD's Facs Lysing solution? Please advice. Thanks, Perla Perla Nunes Dept. of General Surgery Research Cannon Research Center Charlotte, NC 28232 704-355-7269 perla.nunes@carolinashealthcare.org *********************************************************************** This electronic message may contain information that is confidential and/or legally privileged. It is intended only for the use of the individual(s) and entity named as recipients in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from any computer. Do not deliver, distribute or copy this message, and do not disclose its contents or take any action in reliance on the information it contains. Thank you.
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