PMC:2779105 / 193030-194169 JSONTXT

{"project":"2_test","denotations":[{"id":"19956594-19412552-88163132","span":{"begin":114,"end":115},"obj":"19412552"},{"id":"T5796","span":{"begin":114,"end":115},"obj":"19412552"}],"text":"Preparation of Calcium Granules from Serum\nCalcium granules were prepared as described earlier for serum pellets [3]. Briefly, the granules shown in this study (labeled as “Calcium Granules”) were prepared by adding sterile solutions of either 0.25 M CaCl2 and/or 0.25 M Na2HPO4 (both at pH 7.4) to FBS (2.5 ml) at final concentrations of either 48 mM CaCl2 (used for Figs. 9G, 10D, 12D, 13D, 14D, and 15G), 24 mM Na2HPO4 (Figs. 9H, 10E, 12E, 13E, 14E, and 15H), or 2 mM of both CaCl2 and Na2HPO4 (Figs. 9I, 10F, 12F, and 14F). Calcium granules were also prepared in HS (2.5 ml) by diluting 2 mM of both CaCl2 and Na2HPO4 (Figs. 13F and 15I). The ion solutions were filtered through both 0.2-µm and 0.1-µm membranes prior to use and were added in a drop-wise manner with vigorous shaking in order to avoid spontaneous precipitation. Treated sera were incubated at room temperature overnight, followed by centrifugation at 16,000× g for 1 hour, and washing steps using HEPES buffer and the same centrifugation steps. The granules were resuspended in 100 µl of HEPES buffer and were used for the various microscopy and spectroscopy analyses."}