I am performing immunofluorescent staining of rat spleen cells. I am using a stomacher to prepare single cell suspensions and ammonium chloride to lyse RBCs. I have found that the cell viability and antibody binding is affected. I have attempted to analyze without RBC lysing, but I have not been able to eliminate the RBCs electronically. Does anyone have a procedure for CD45 antibody labeling of rat spleen cells for gating of lymphocyte subsets and excluding RBCs? I am also interested in other procedures for processing rat spleen, RBC lysing, or flow analysis without lysing. Thanks! Judy Miller Merck & Co., Inc. Phone: 215-652-3127 Fax: 215-652-7758 E-mail: millerj@merck.com
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