Alexa Fluor® 647 Mouse Anti-Human CD24

Protocol tips

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		Fluorescence-activated cell sorting BD, FACSAria II SORP, and FACSDiva 8.0.1 software were used. After sorting, cells were seeded in ultra-low adherent plates for tumorsphere formation assays and treated with SFN or control (0.9% NaCl) for 7–10 days.

Downstream tips
Fluorescence-activated cell sorting BD, FACSAria II SORP, and FACSDiva 8.0.1 software were used. After sorting, cells were seeded in ultra-low adherent plates for tumorsphere formation assays and treated with SFN or control (0.9% NaCl) for 7–10 days.

Publication protocol

MDA-MB-231-luc-D3H1 cells were stained with the following antibodies: FITC Rat Anti-human CD49f, PE Mouse Anti-human CD44, Alexa fluor 647 Mouse Anti-human CD24 (BD Pharmingen™). Fluorescence-activated cell sorting BD, FACSAria II SORP, and FACSDiva 8.0.1 software were used. Stained cells were examined and sorted for stem/progenitor cells with CD49f+ population followed by a final gate to select CD44+/CD24- within the CD49+ cell fraction. A total of 1.39x106 cells were sorted. The percentage of parent population were 98.31 of CD49f+ marker, followed by a sequential gate of CD24+ and CD44+ with 0.01% and 1.30% of parental population respectively. After sorting, cells were seeded in ultra-low adherent plates for tumorsphere formation assays and treated with SFN or control (0.9% NaCl) for 7–10 days.

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