Cells were collected, stained, and analyzed by flow cytometry (BD Cytoflex S flow cytometer) as follows. Staining for membrane surface molecules included use of the following antibodies: EpCAM-PE (phycoerythrin) : [VU-1D9] (Abcam) and isotype control [MOPC-21] (BD Pharmingen); E-cadherin-PE : CD324 [DECMA-1] BioLegend) and isotype control [A95-1] (BD Pharmingen); and N-cadherin-APC (Allophycocyanin) : CD325 [8C11] (BioLegend) and isotype control [27 (link)–35 (link)] (BD Pharmingen). Staining was performed following standard methods. Cells were stained for 30 min (EpCAM) and 20 min (E-cadherin and N-cadherin). For intracellular staining of CCT2 we used the antibody CCT2-PE : [NP_006422] (LSBio) and the isotype control [MOPC-21] (LSBio), following the method in ThermoFisher Scientific’s “Protocol A: two-step protocol: intracellular (cytoplasmic) proteins” and incubating the antibody for 70 min. When optimizing the CCT2 intracellular stain for CSS Autoprep conditions, we adjusted the antibody staining protocol for CCT2-PE to match the 20 min in the CSS Autoprep automated conditions. All data was generated using FCS Express 6 software (De Novo).
Epcam pe
Manufactured by Abcam
Sourced in United Kingdom
EpCAM-PE is a lab equipment product that detects the presence of the epithelial cell adhesion molecule (EpCAM) in samples. EpCAM is a transmembrane glycoprotein that is expressed on the surface of epithelial cells. The EpCAM-PE product utilizes a phycoerythrin (PE) fluorescent label to enable the visualization and quantification of EpCAM-positive cells.
Automatically generated - may contain errors
2 protocols using epcam pe
1
Flow Cytometric Analysis of EMT Markers
2
Multiparametric Flow Cytometry Profiling
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EECs were blocked with 10% normal goat serum for 10 min on ice. Cells were then either left unstained (negative control) or incubated with mouse anti-human E-cadherin-BV421 (BD Biosciences, Oxford, UK), mouse anti-human epithelial cell adhesion molecule (EpCAM)-PE (Abcam, Cambridge, UK), mouse anti-human CD31-PerCP-Cy5.5, and rat anti-human CD45-APC-Cy7 (BioLegend, London, UK). Flow cytometry was carried out on a BD LSR Fortessa 5 L flow cytometer (BD Biosciences, Oxford, UK). Analysis was carried using FlowJo software (BD Biosciences, Oxford, UK).
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