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Fluorescence signal and dual parameter plots from Alexa Fluor 594 Click-iT Plus EdU Flow Cytometry Assay Kit, CD3-APC, and FxCycle Violet
Jurkat (human T-cell leukemia) cells were treated with 10 µM EdU for 2 hours, stained with CD3-APC (Cat. No MHCD0305) and detected according to the recommended staining protocol. The figures show a clear separation of proliferating cells which have incorporated EdU and non-proliferating cells which have not. Panel A shows data from cells labeled with Alexa Fluor™ 594 picolyl azide analyzed on a BD™ LSRII flow cytometer using 532 nm excitation and a 585/42 nm bandpass emission filter; Panel B shows the same cells using 633 nm excitation and a 660/20 nm bandpass emission filter for detection of the CD3-APC and 532 nm excitation and a 585/42 nm bandpass emission filter for detection of the Alexa Fluor 594 picolyl azide; Panel C shows the dual parameter plot of the Click-iT™ Plus EdU Alexa Fluor™ 594 and FxCycle™ Violet. Data were collected and analyzed using a BD LSRII flow cytometer using 532 nm excitation and a 585/42 nm bandpass emission filter for detection of the Alexa Fluor 594 picolyl azide and 405 nm excitation and a 450/50 nm bandpass emission filter for detection of the FxCycle Violet fluorescence. This figure combines DNA content with EdU; cells that are positive for both labels are in S-phase of the cell cycle.
