You can enrich normal reticulocytes using linear BSA gradients (1). In any event, erythrocytes and presumably their precursors require serum albumin as a membrane stabilizer. Even small differences in tonicity will effect the buoyant density of cells. (1) R. C. Leif and J. Vinograd; “The Distribution of Buoyant Density of Human Erythrocytes in Bovine Albumin Solutions”. Proc. N.A.S. 51, pp. 520-528 (1964). -----Original Message----- From: Dana Levasseur [mailto:Dana_Levasseur@microbio.uab.edu] Sent: Wednesday, November 10, 1999 8:25 AM To: cyto-inbox Subject: Reticulocyte purification/enrichment Hello All, We're working with several different transgenic knockout lines of mice which exhibit a severe peripheral blood reticulocytosis as well as hypercellular bone marrow. It would be nice to find a reliable enrichment or purification protocol to look at retics and stress retics in these animals to verify the numbers I am seeing with thiazole orange and other nucleic acid-staining dyes like SYTO. The literature mentions exploiting ion gradient characteristics but I want to try to keep the mouse cells in a reasonably physiological buffer as they are a little less hardy than human RBCS. Anyone have any experience or ideas? Much appreciated! Dana Dana Levasseur Dept. of Biochemistry and Molecular Genetics University of Alabama at Birmingham 845 19th Street South Bevill Biomedical Research Building Rm. 870 Birmingham, AL 35294 Tel: 205-933-5334 Email: dnl@uab.edu
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