Alexa Fluor® 488 Click-iT® Plus EdU Flow Cytometry Assay Kits, mCherry, and FxCycle™ FarRed

A549 cells were transduced with adenovirus for 48 hours, treated with 10 µM EdU for 2 hours and detected according to the recommended staining protocol. The figures show a clear separation of proliferating cells that have incorporated EdU and nonproliferating cells that have not. Panel A shows data from cells labeled with Alexa Fluor® 488 picolyl azide on an LSRII flow cytometer using 488 nm excitation and a 530/30 nm bandpass emission filter. Panel B shows the mCherry-expressing cells clicked with Click-iT® Plus EdU Alexa Fluor® 488 picolyl azide using 532 nm excitation and a 585/42 nm bandpass emission filter to display the mCherry fluorescence. Panel C shows the mCherry-expressing control cells clicked with Click-iT® Plus EdU Alexa Fluor® 488 picolyl azide but without the copper in the reaction, using 532 nm excitation and a 585/42 nm bandpass emission filter to display the positive control mCherry fluorescence. Panel D shows the dual parameter plot of the Click-iT® Plus EdU Alexa Fluor® 488 and FxCycle™ FarRed. Data were collected and analyzed using an LSRII flow cytometer using 488 nm excitation and a 530/30 nm bandpass emission filter for detection of the EdU Alexa Fluor® 488 picolyl azide and 635 nm excitation and a 660/20 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.

A549 cells were transduced with adenovirus for 48 hours, treated with 10 µM EdU for 2 hours and detected according to the recommended staining protocol. The figures show a clear separation of proliferating cells that have incorporated EdU and nonproliferating cells that have not. Panel A shows data from cells labeled with Alexa Fluor® 488 picolyl azide on an LSRII flow cytometer using 488 nm excitation and a 530/30 nm bandpass emission filter. Panel B shows the mCherry-expressing cells clicked with Click-iT® Plus EdU Alexa Fluor® 488 picolyl azide using 532 nm excitation and a 585/42 nm bandpass emission filter to display the mCherry fluorescence. Panel C shows the mCherry-expressing control cells clicked with Click-iT® Plus EdU Alexa Fluor® 488 picolyl azide but without the copper in the reaction, using 532 nm excitation and a 585/42 nm bandpass emission filter to display the positive control mCherry fluorescence. Panel D shows the dual parameter plot of the Click-iT® Plus EdU Alexa Fluor® 488 and FxCycle™ FarRed. Data were collected and analyzed using an LSRII flow cytometer using 488 nm excitation and a 530/30 nm bandpass emission filter for detection of the EdU Alexa Fluor® 488 picolyl azide and 635 nm excitation and a 660/20 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.

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