Single Cell Surface Marker Isolation

Single cells were suspended in PBS, 2 mM EDTA, 0.5% BSA and stained with antibody for 15
min at 4°C. Fluorescence-activated cell sorting and analysis were performed on a BD Special
Order FACS Aria II system and Diva v6.1.1 (BD Biosciences). Live single cells were gated based on
scatter properties and analyzed for their surface marker expression as previously described
[15 (link)]. Antibodies used for FACS include Epcam-PE
(Miltenyi Biotech), CD44⁻FITC (ebioscience), and CD49f-APC (BioLegend).

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Liu H., Cadaneanu R.M., Lai K., Zhang B., Huo L., An D.S., Li X., Lewis M.S, & Garraway I.P. (2015). Differential Gene Expression Profiling of Functionally and Developmentally Distinct Human Prostate Epithelial Populations. The Prostate, 75(7), 764-776.

Publication 2015

BD SpecialOrder FACS Aria II system Diva v6.1.1 Epcam-PE CD44−FITC CD49f-APC

Antibodies Antibody Aria Cd44 Cd49f Cells Edta Epcam Fitc

Corresponding Organization : VA Greater Los Angeles Healthcare System

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Variable analysis

independent variables

Antibody staining

dependent variables

Surface marker expression

control variables

Cell suspension in PBS, 2 mM EDTA, 0.5% BSA
Staining duration of 15 min at 4°C
Fluorescence-activated cell sorting and analysis performed on a BD Special Order FACS Aria II system and Diva v6.1.1 (BD Biosciences)
Gating of live single cells based on scatter properties

controls

No positive or negative controls were explicitly mentioned in the provided information.

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