FITC BrdU Flow Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cells were treated with BrdU (10 μM) for 4 hours.	- Cells were treated with APC-conjugated anti-BrdU antibody

Upstream tips
- Cells were treated with BrdU (10 μM) for 4 hours.
Protocol tips
- Cells were treated with APC-conjugated anti-BrdU antibody

Publication protocol

RAW 264.7 cells exposed to NPs or LPS for variable times were washed, counted, and plated in fresh media at 105 cells/mL. Cells were pulsed with BrdU (10 μM) for 4 hours, after which cells were processed as recommended by the vendor. Briefly, cells were fixed, permeabilized, and stained with APC-conjugated anti-BrdU antibody. Cell cycle status was determined by analyzing BrdU signal versus 7AAD or SYTO® Green (for Au NPs-treated cells) staining using DIVA software. Cell cycle status is expressed as cell percentage. In order to detect possible interference of NPs with the assay, control cells were exposed to BrdU and NPs simultaneously and their MFI compared to control cells only exposed to BrdU (ie, a spiked control).

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