1mM EDTA is definitely fine. Have not tested higher concentrations ======================================================= Rachel M. Gerstein, Ph.D. Associate Professor Department of Molecular Genetics and Microbiology Graduate Program in Immunology/Virology University of Massachusetts Medical School 55 Lake Avenue North Worcester, MA 01655-0002 (508) 856-1044 (508) 856-5920 (FAX) -----Original Message----- From: Karis Hughes [mailto:karismhughes@yahoo.com] Sent: Thu 3/20/2008 4:16 PM To: cyto-inbox Subject: 1mM EDTA FACS buffer and extensive fluorochrome panel Hi Flow~ers- I have a practical question that someone might easily be able to answer. We have been using a 10 color panel with the LSRII with great luck in analysis of non adherent cells. For this purpose we usually do not have EDTA added to our FACS buffer. However, we have recently been interested in some adherent cell populations. We have been running our samples over mesh to get rid of clumps, then adding 25ul of 5mM EDTA to our 200ul sample immediately prior to running them. However we are still running in to small clumping issues. Here's the question: Does anyone know if the fluorochromes APC, APC-Cy7, PE-Cy7, PE-Cy5.5, Alexa700, PacBlu, Q-Dot are stable in 1mM EDTA or 5mM EDTA supplemented FACS buffer for a small period of time such as a few hours? Thanks so much for sharing your experience- Karis Hughes Graduate Student Department of Immunology UT-Southwestern Medical Center Karis M. Hughes karismhughes@yahoo.com Home: 972-444-0198 Fax: 952-516-1457Received on Mon Mar 24 15:18:00 2008
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