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- W112517859 abstract "Publisher Summary This chapter discusses some technical considerations for the separation of human peripheral blood lymphocyte subpopulations by rosetting with sheep erythrocyte. There are several points in the E-rosette forming cells (E-RFC) procedure where difficulties may be encountered. Best results are obtained when platlets are removed by aggregation with ADP prior to FH separation of peripheral blood mononuclear leukocytes (PBL) from whole blood. For optimal E-RFC formation, the sheep erythrocytes should be fresh (never older than two weeks) and from a sheep selected to form strong E rosettes. Fetal calf serum (FCS) employed in rosetting should be selected from a lot giving maximum numbers of E-RFC and should be absorbed with E if there is evidence of agglutinins. For rosette formation, PBL and E are usually pelleted together by centrifugation, and the optimum E:PBL ratio is approximately 40:1. A round-bottom centrifuge tube and the use of cold (4°C) media run down the tube's side will allow easier resuspension. The procedure for E-RFC formation was simplified by eliminating the centrifugation step. With this procedure, a higher E:PBL ratio of 80:1 and a final concentration of 40% fetal calf serum is used." @default.
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- W112517859 date "1977-01-01" @default.
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- W112517859 title "TECHNICAL CONSIDERATIONS FOR THE SEPARATION OF HUMAN PERIPHERAL BLOOD LYMPHOCYTE SUBPOPULATIONS BY ROSETTING WITH SHEEP ERYTHROCYTE" @default.
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- W112517859 doi "https://doi.org/10.1016/b978-0-12-458050-3.50099-0" @default.
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