Cell pellets from tissue dissociation were resuspended in 100 μl of FACS stain buffer (2% FBS in PBS) with 1 μl of anti–CD31–Brilliant Violet 786 (clone 390, BD Biosciences), 1 μl of anti–CD45–APC/Cy7 (clone 30-F11, BioLegend), 1 μl of parenchymal cell marker [anti–GLUT2-PE (Bioss), anti–EPCAM-PE (BioLegend), or anti–cTnT-PE (Miltenyi Biotec)], 0.02 μl of Alexa Fluor 647–conjugated AxV, 5 ml Zombie Violet (BioLegend), and 0.1 μl of CellEvent Caspase-3/7 Green Detection Reagent (Thermo Fisher Scientific). Cells were stained on ice for 45 min away from the light and then centrifuged at 200g for 5 min at 4°C, and fluorescence in different cell types was analyzed on an Attune NxT flow cytometer (Thermo Fisher Scientific).
Anti epcam pe
Manufactured by BioLegend
Sourced in United States
Anti–EPCAM-PE is a fluorescently-labeled antibody that specifically binds to the EPCAM protein. It can be used for the detection and analysis of EPCAM-expressing cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Cellevent caspase 3 7 green detection reagent, by Thermo Fisher Scientific (1 mentions)
Anti cd45 apc cy7 clone 30 f11, by BioLegend (1 mentions)
Attune nxt flow cytometer, by Thermo Fisher Scientific (1 mentions)
Zombie violet, by BioLegend (1 mentions)
Facscalibur, by BD (1 mentions)
Lamina propria dissociation kit, by Miltenyi Biotec (1 mentions)
3 protocols using anti epcam pe
1
Multicolor Flow Cytometry of Tissue Cells
2
Flow Cytometry Cell Staining Protocol
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Flow cytometry cell staining was performed at 4°C in PBS supplemented with 2% FBS unless
otherwise indicated. The antibodies used for flow cytometry and functional studies
included human EpCAM-biotin (Acro, Beijing, China), anti-biotin-APC (Biolegend, San Diego,
CA, USA), anti-EpCAM-PE (Biolegend). Data were analyzed using NovaExpr software (FlowJo,
LLC, Ashland, OR, USA).
otherwise indicated. The antibodies used for flow cytometry and functional studies
included human EpCAM-biotin (Acro, Beijing, China), anti-biotin-APC (Biolegend, San Diego,
CA, USA), anti-EpCAM-PE (Biolegend). Data were analyzed using NovaExpr software (FlowJo,
LLC, Ashland, OR, USA).
3
Isolation and Purification of Intestinal Epithelial Cells
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IECs were isolated from intestinal tissue using the Lamina Propria Dissociation Kit (Miltenyi BioTech, Bergisch Gladbach, Germany) according to the manufacturer’s protocol with minor deviations as described before [19 (link)]. In brief, intestinal epithelial cells were isolated by disruption of the structural integrity of the epithelium using ethylenediaminetetraacetic acid (EDTA) and dithiothreitol (DTT). The purity of individual IEC fractions was analyzed by flow cytometry on a FACS Calibur flow cytometer (B&D, Heidelberg, Germany) with Cellquest analysis software version 5.1 (B&D, Heidelberg, Germany). We used the Anti-EpCam-PE (Clone: G8.8, Biolegend, San Diego, CA, USA) antibody for analysis of IEC purity. FACS data was analyzed using Flowing Software version 2.5.1 (Perttu Terho, Turku Centre for Biotechnology, Finland).
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