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2 protocols using anti cd66abce fitc

1

Multi-Marker Flow Cytometry Profiling

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Antibodies anti-CD31 PE-Cy7, anti-CD18 PE or APC, anti-CD51 FITC, anti-CD64 FITC, anti-CD14 FITC, PE or PerCP-Cy5.5, anti-CD66abce FITC, anti-CD34 VioBlue®, anti-CD38 FITC, anti-CD44 PE, anti-CD45 VioBlue® or APC-Vio770, anti-CD3 FITC or PerCP-Cy5.5, anti-CD20 FITC or PerCP-Cy5.5, anti-CD15 FITC, anti-CD16 PerCP-Cy5.5, anti-HLA-DR VioBlue®, anti-CD86 VioBlue®, anti-CD80 VioBlue®, anti-CD90 FITC and PerCP-Cy5.5, anti-CD133 APC, anti-CD138 PE, anti-CD140a PE, anti-CD146 FITC, and anti-MSCA-1 PE were from Miltenyi Biotec. Anti-STRO-1 AlexaFluor® 647, anti-CD73 PE-Cy7, anti-CD105 PerCP-Cy5.5, anti-CD140b PE, anti-CD56 APC, anti-CD309 PerCP-Cy5.5, anti-CD115, anti-CD144 PE, anti-SSEA-3 FITC, and anti-SSEA-4 FITC were from BioLegend (San Diego, CA). Anti-CD271 PE, anti-CD11b PE, anti-CD11c PE, and anti-CD36 APC were from BD Biosciences (San Jose, CA).
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2

Multiparametric Flow Cytometry Immunophenotyping

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Freshly isolated MPCs and differentiated MSCs were processed for flow cytometry analysis, incubating cells for 30 min at 4°C with a set of fluorochrome-conjugated antibodies, including: anti-CD31 PE-Cy7, anti-CD18 APC, anti-CD14 FITC, anti-CD66abce FITC, anti-CD34 VioBlue, anti-CD90 FITC, anti-CD146 FITC, and anti-MSCA-1 PE were from Miltenyi Biotec. Anti-STRO-1 AlexaFluor 647, anti-CD73 PE-Cy7, and anti-CD105 PerCP-Cy5.5 were from BioLegend. The anti-CD11c PE antibody was from BD Bioscience. After cell washing with MACSQuant Running Buffer (Miltenyi Biotec), samples were acquired on MACSQuant Flow Cytometer (Miltenyi Biotec) and analyzed by MACSQuantify Analysis Software (Miltenyi Biotec). Proper isotype controls (all from Miltenyi Biotec) were also employed to set the positive regions of the histograms.
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