Flow cytometric analysis was performed to identify the ST cells expressing Mincle [13 (link)]. ST were digested with α-minimum essential medium (α-MEM, Invitrogen, CA, USA) containing 1 mg/ml collagenase solution (Sigma, MO, USA) on shaker for 2 hours at 37°C. Extracted cells were incubated with phycoerythrin (PE)-conjugated mouse anti-CD14 monoclonal antibody (1 : 20; clone M5E2, Biolegend, CA, USA), fluorescein isothiocyanate (FITC)-conjugated mouse anti-CD45 monoclonal antibody (1 : 20; clone LCA, BD Biosciences, San Jose, CA, USA), allophycocyanin (APC)/Cy7-conjugated anti-human CD31 (1 : 20; clone WM59, Biolegend), APC-conjugated mouse anti-CD90 monoclonal antibody (1 : 20; clone 5E10, Biolegend), and rat anti-Mincle monoclonal antibody (1 : 200; clone 1H2, MBL, Nagoya, Japan) for 45 minutes at 4°C. The cells were washed twice in PBS, resuspended in ice cold PBS and reacted with brilliant violet 421-conjugated goat anti-rat IgG polyclonal antibody (1 : 100; clone Polu4054, Biolegend). After washing twice in PBS, the cells were analyzed by flow cytometric device (FACSVerseTM; BD Biosciences). Isotype controls were used as the posi-tive gate.
Apc cy7 conjugated anti human cd31
Manufactured by BioLegend
Sourced in United States
The APC)/Cy7-conjugated anti-human CD31 is a lab equipment product that binds to the CD31 receptor, also known as PECAM-1, on the surface of human cells. It is labeled with the APC/Cy7 fluorescent dye, which can be used for flow cytometry and other applications.
Automatically generated - may contain errors
Lab products found in correlation
α mem, by Thermo Fisher Scientific (1 mentions)
Facsverse, by BD (1 mentions)
Collagenase solution, by Merck Group (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Accumax, by Merck Group (1 mentions)
Accumax solution, by Merck Group (1 mentions)
Apc conjugated cd105, by BioLegend (1 mentions)
2 protocols using apc cy7 conjugated anti human cd31
1
Flow Cytometric Analysis of Mincle+ ST Cells
2
Flow Cytometry Analysis of Lung Cell Populations
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3D SAEC-F11 VEGFhigh- HMVEC-L and SAEC-F11-HMVEC-L lung aggregates were cultured for 72 h in the presence or absence of rhWnt5a. Aggregates were then dissociated with AccumaxTM (Sigma-Aldrich, St. Louis, USA) solution and washed in PBS once. Single cell suspensions were incubated with Allophycocyanin (APC) conjugated anti-human CD105 (Clone 43A3, BioLegend, San Diego, USA) and Brilliant Violet 421 conjugated anti-human CD31 (Clone VM59, BioLegend, San Diego, USA) for 30min at room temperature in dark. Native lung AC and SCC samples were dissociated by enzymatic digestion (Accumax Solution, Sigma Aldrich, St. Louis, USA) and the single cell suspensions were washed in PBS once, then cells were incubated with APC Cy7 conjugated anti-human CD31 (Clone VM59, BioLegend, San Diego, USA) and APC conjugated CD105 (Clone 43A3, BioLegend, San Diego, USA) antibodies. Cells then were washed in PBS, fixed with 1% PFA and stored at 4 °C in dark until FACS analysis. Labeled cells were analyzed using FACS Canto II flow cytometer (BD Immunocytometry Systems, Erembodegen, Belgium) with BD FACS DIVA software V6 and data were analyzed by FCS Express V3 software.
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