The morning of the copulation plug was designated embryonic day (E) 0.5 and embryos were harvested from pregnant dams on 11.5, E12.5, E14.5, and E16.5 into ice-cold PBS. The small intestine (digestive tract distal to the pylorus and proximal to the cecum) was digested with 0.25% trypsin (Life Technologies) for 30 min at 37°C to release single cells, followed by neutralization with fetal bovine serum (FBS, Life Technologies). Cells were passed over 40-μm filters to remove tissue fragments, centrifuged at 1,200 g for 5 min, washed in cold PBS, suspended in fluorescent-activated cell sorting (FACS) buffer (5 mM EDTA in PBS and 2% FBS), stained with APC-conjugated EpCAM antibody (Biolegend 118214, Lot B217174, 1:100) for 1 h at 4°C. Viable (DAPI−) EpCAM+ cells were isolated by flow cytometry on a FACS Aria II SORP instrument.
Apc conjugated epcam antibody
Manufactured by BioLegend
Sourced in China
The APC-conjugated EpCAM antibody is a laboratory tool used for the detection and analysis of EpCAM (Epithelial Cell Adhesion Molecule) expression on cells. The antibody is conjugated with the fluorescent dye Allophycocyanin (APC), enabling the visualization and quantification of EpCAM-positive cells by flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by BioLegend (1 mentions)
Moflo xdp flow cytometer, by Beckman Coulter (1 mentions)
Propidium iodide, by BioLegend (1 mentions)
Cd24 antibody, by BioLegend (1 mentions)
Epcam, by BioLegend (1 mentions)
2 protocols using apc conjugated epcam antibody
1
Isolation of Epithelial Cells from Embryonic Intestine
2
Quantifying Liver Cancer Stem Cells
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To analyze the proportion of epithelial cell adhesion molecule (EpCAM) or CD24-positive liver CSCs, the colorectal cells were harvested and resuspended in the staining buffer, and then incubated with allophycocyanin (APC)-conjugated EpCAM antibody (BioLegend, Shanghai, China) or CD24 antibody (BioLegend) for 30 minutes at 4 ℃ in the dark. The cells were further washed with cold staining buffer twice and resuspended in the staining buffer containing 1 µg/mL propidium iodide (PI; BioLegend) followed by flow cytometry analysis using a Moflo XDP flow cytometer from Beckman Coulter, CA, USA. For CD24+ or EpCAM+ cell sorting, the colorectal cells (5×107) were harvested and resuspended in cold staining buffer, then incubated with antibodies against human CD24 (BioLegend) or EpCAM (BioLegend), respectively. Positively and negatively stained cells were then sorted using the Moflo XDP flow cytometer. The sorted cells from three independent experiments were subjected to a real-time polymerase chain reaction (PCR) assay.
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