The Amplex Red assay can be used to determine cellular cholesterol levels in AML cells.
(A) The Amplex Red assay yields cholesterol measurements that are linear across at least 1 to 16 μM purified cholesterol input when measured 60 to 120 minutes after reaction initiation. afu indicates arbitrary fluorescence units. (B) Total cell cholesterol content can be measured when lysates from 2.5 × 104 to 10 × 104 NB4 cells in 100 μL assay volume are used as the cholesterol input. (C) Lysates from 105 NB4, HL60, or KG1a cells in 100 μL assay volumes yield Amplex Red assay measurements of 4.4, 3.6, and 3.3 μM total cholesterol per assay, respectively, equivalent to 4.4, 3.6, and 3.3 nmol cholesterol per 106 cells. Data are presented with standard errors and represent the mean from 8 to 20 independent replicates.
