Passage 1 cultures were dissociated by treatment with Accutase for 10 min at 37°C. Cell suspensions were passed through 40 µm cell strainers into 4× volume of DMEM/F-12 and centrifuged for 5 min, 200× g. Approximately 5 × 105 cells per replicate were resuspended in MACS buffer and incubated with the CD31-APC antibody (Miltenyi Biotec) for 30 min at 4°C, protected from light. Cell suspensions were brought to a volume of 5 mL with MACS buffer and centrifuged at 4°C for 5 min, 200× g. Cells were resuspended in 500 µL MACS buffer containing 2 µg/mL DAPI and 0.5 µL Vybrant DyeCycle Green Stain (Invitrogen) and incubated at RT for 1 hr, protected from light. Cells were analyzed on an Attune NxT flow cytometer (Invitrogen). FlowJo software (BD Biosciences) was used to gate CD31+ cells and quantify the percentage of S/G2/M phase cells.
Cd31 apc antibody
Manufactured by Miltenyi Biotec
The CD31-APC antibody is a laboratory reagent used for the detection and analysis of CD31-positive cells in various biological samples. CD31, also known as PECAM-1, is a cell surface glycoprotein expressed on endothelial cells, platelets, and some immune cells. The APC (Allophycocyanin) fluorescent label allows for the identification and quantification of CD31-positive cells using flow cytometry or other cell-based analysis techniques.
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2 protocols using cd31 apc antibody
1
Quantification of Endothelial Cells
2
Immune cell isolation and analysis
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Cells were first incubated with mouse BD FC Block (BD Biosciences, #553141), to block nonantigen-specific binding of immunoglobulins in a dilution of 1:50. Endothelial and hematopoietic cells were stained directly after isolation with CD45-PE antibody (Miltenyi, #130-102-781) or CD31-APC antibody (Miltenyi, #130-102-978) according to manufacturer's protocol. Purity of cells was quantified using FACS Canto II devices (BD Bioscience).
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