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To assess the ability of anti-CD73 antibodies to inhibit CD73 activity, cells were incubated with antibodies prior to addition of 250 mM AMP. Phosphate levels were measured in the cell culture supernatant using the Sensolyte Malachite Green assay kit (AnaSpec, Cat# AS-71103). CD73 activity was also measured using the CellTiterGlo assay kit (Promega, Cat#G7571) as previously described.21 22 A cell free reaction was used to determine maximum inhibition of the CellTiterGlo reaction by AMP.
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