Binding sites for angiotensin II in human mononuclear leucocytes

K Shimada, Y Yazaki - The Journal of Biochemistry, 1978 - jstage.jst.go.jp
K Shimada, Y Yazaki
The Journal of Biochemistry, 1978jstage.jst.go.jp
Preparation of Cells: The blood cells from normal adults were fractionated on Ficoll-
Isopaque gradients according to Boyum(8). The isolated mononuclear leucocytes were
resuspended to a final concentration of (3-8) x 107 cells/ml in a Tris-HCl buffer (pH 7.4)
containing 120 mm NaCl, 20 mm Tris, 5 mm dithiothreitol, 10-4 M phenyl methylsulfonyl
fluoride, and 5 mm EDTA. The isolated leucocytes consisted of approximately 99 of
mononuclear leucocytes and 1% of granu locytes. Erythrocytes and platelets comprised 20 …
Preparation of Cells: The blood cells from normal adults were fractionated on Ficoll-Isopaque gradients according to Boyum(8). The isolated mononuclear leucocytes were resuspended to a final concentration of (3-8) x 107 cells/ml in a Tris-HCl buffer (pH 7.4) containing 120 mm NaCl, 20 mm Tris, 5 mm dithiothreitol, 10-4 M phenyl methylsulfonyl fluoride, and 5 mm EDTA. The isolated leucocytes consisted of approximately 99 of mononuclear leucocytes and 1% of granu locytes. Erythrocytes and platelets comprised 20 and 15% of the total number of cells in suspension respectively.
Binding Studies: 50 jut of cell suspension and [1251] angiotensin‡ U(208 ƒÊM) were added to an incubation medium of Krebs-Ringer-phosphate buffer(pH 7.4) containing 0.2% bovine serum albumin and 5 mm EDTA in a final volume of 0.2 ml. Incubations were performed at 22 C in siliconized test tubes both in the absence and presence of unlabeled angiotensin‡ U(4). At the end of the incubation period, 2 ml of ice-cold
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