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You can avoid bruising by alternating the sites

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Successively the cells were stimulated overnight with TNF- (100 ng/mL), washed with Dulbecco's PBS added with 2% BSA (Bovine Serum Albumine, fraction V, Sigma-Aldrich) and CaCl 2 -MgCl 2 0,7 mM (Sigma-Aldrich), and then incubated with mouse mAb anti-human VCAM1 (Sigma-Aldrich) 5 g/mL for 90 min at RT