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Assay Method Information |
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| | Exogenous PldA Assay |
| Description: | In brief, 6 nM purified PldA was incubated with liposomes containing 90 μM 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol), 10 μM 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, and 2.5 μCi [3H]-DPPC for 10 min at 37 °C in buffer containing 50 mM HEPES, pH 7.5, 100 mM KCl, and 3 mM MgCl2. The reaction was quenched with 10% trichloroacetic acid and bovine serum albumin on ice. Protein and lipid were removed by centrifugation. Free [3H]-choline was measured by scintillation counting. |
| Affinity data for this assay | |
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