From: mattiaqu@tin.it
Date: Thu Dec 18 2003 - 04:32:58 EST
Dear Professor Sutherland, I would like to thank you for your kind mail. I'm actually interested in using the boolean gating strategy to identify CD90+ and c-Kit+ stem cells directly, which is to say possibly all CD90+ or c-kit positive cells, even those that don't belong to CD34+ subpopulations. I must confess that for the first experiments I did not follow properly the ISHAGE procedure because I've been using a CD45-PerCp antibody and a CD34-biotin conjugated antibody (using a streptaividin-Alexa488 as secondary). So I'm using the brightest dye for the most represented antigen. The result therefore are not reliable so far. I will switch the dyes for the next experiments, but I was thinking to keep using the Per-Cp dye instead of using PE. May I ask you your opinion on using primary antibodies conjugated with biotin having streptavidin-dye as secondary for this kind of evaluations? I will be glad to send you a sample listmode file and actually I've been trying to send it directly to your e-mail adress but I think that my mails have been rejected. Best regards, Mattia MATTIA QUARTA M.D. Department of Medical and Surgical Sciencies University of Padua Via Giustiniani 2 35100, PADOVA Italy Phone: +39-049-8211873 fax: +39-049-8211884
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