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Flow cytometric analysis of the conversion of BD Pharmingen™ DAF-FM DA to fluorescent DAF-FM by nitric oxide in Jurkat Cells treated with DEA NONOate. Jurkat cells in 1× DPBS were preincubated with (solid line) or without (dashed line) 10 µM BD Pharmingen™ DAF-FM DA for 30 minutes at 37°C and then treated with (red) or without (blue) 10 µM Diethylamine NONOate (Sigma Aldrich, Cat. No. D5431) for 30 minutes at 37°C. Cells were washed once and then analyzed by flow cytometry. Treatment with the nitric oxide donor Diethylamine NONOate results in an increase in the fluorescence of DAF-FM DA loaded cells. Histograms showing the levels of the nitric oxide-induced fluorescent benzotriazole derivative were derived from gated events with the light scattering characteristics of intact Jurkat cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
BD Pharmingen™ DAF-FM DA
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