Hi, How can I block non-specific binding in procedure of purification B cells from PBMNC using negative immunoselection by immunomagnetic beads. For that purpose, I tried to use different concentrations of human Ig (to block Fc receptors), but still, I have got only 5-6 million of B cells from 200 million PBMN cells because of non-specific binding. By the way for this purpose I use anti-CD3, CD5, CD56, CD16, CD14 CD13 and anti-glycophorin. My next question is about ApoAlert mitochondrial detection kit by flow-cytometry (Clonitech company), and MitoSensor, dye that company gives in this kit for mitochondrial staining, that is used for determination of “healthy” mitochondria and mitochondria that are changed in the process of apoptosis. I would like to know what is the experience of people that use this kit, because the company provide only technical support for this kit, but not the examples of results that were got. Sincerely, Bratislav Janjic, B.Sc. Department of Pathology E1005 Biomedical Science Tower University of Pittsburgh Cancer Institute PA 15213 __________________________________________________________________________ Visit http://www.visto.com/info, your free web-based communications center. Visto.com. Life on the Dot.
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