The characterization of ECFCs was performed using specific antibodies conjugated to fluorochromes (fluorescein isothiocyanate [FITC], phycoerythrin [PE] and peridin chlorophyll protein [PerCP]), which detect the following specific endothelial surface markers: anti‐CD31‐FITC, clone MBC 78.2 (Invitrogen); anti‐CD144‐PE, clone TEA 1/31 (Beckman Coulter); anti‐CD146PE, clone 128,018 (R&D Systems); anti‐VEGF R2/KDR‐PE, clone 89,106 (R&D Systems); anti‐CD34‐FITC, My10 clone (BD); anti‐CD45‐PerCP, clone 2D1 (BD); anti‐CD133‐APC, clone AC133 (Miltenyi Biotech). The tubes were incubated for 30 min at 4°C, protected from light, washed with PBS, centrifuged at 450 g for 5 min (RT) and resuspended in 300 μl of PBS for the acquisition of 10,000 events on a flow cytometer (FACS Calibur, Immunofluorometry Systems). Data analysis was performed using the BD FACS DIVA software (v.7.0; San Jose, CA, USA). The cells were considered ECFCs if they tested positive for CD31, CD144, CD146 and KDR markers, negative for CD45 and CD133, and exhibited decreased CD34 expression.
Anti vegf r2 kdr pe clone 89 106
Manufactured by R&D Systems
Sourced in United States
Anti‐VEGF R2/KDR‐PE, clone 89,106 is a laboratory reagent used for the detection and analysis of VEGF receptor 2 (KDR) expression on cells. It is conjugated with the fluorescent dye phycoerythrin (PE) for easy detection and quantification by flow cytometry or other compatible techniques.
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2 protocols using anti vegf r2 kdr pe clone 89 106
1
Endothelial Progenitor Cell Characterization
2
Phenotypic Characterization of Endothelial Colony-Forming Cells
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Characterization of ECFC was performed using speci c antibodies conjugated to uorochromes [ uorescein isothiocyanate (FITC), phycoerythrin (PE), peridin chlorophyll protein (PerCP)] that detect speci c endothelial surface markers: anti-CD31-FITC, clone MBC 78.2 (Invitrogen, Camarillo, CA, USA), anti-CD144-PE, clone TEA 1/31 (Beckman Coulter, Marseille, France), anti-CD146PE, clone 128018 (R&D Systems, Minneapolis, MN, USA), anti-VEGF R2/KDR-PE, clone 89106 (R&D Systems, Minneapolis, MN, USA), anti-CD34-FITC, My10 clone (BD, San Jose, CA, USA), anti-CD45-PerCP, clone 2D1 (BD, San Jose, CA, USA), and anti-CD133-APC, clone AC133 (Miltenyi Biotech, Auburn, CA, USA). The cytometric tubes were incubated for 30 min at 4 °C, protected from light, washed with PBS, centrifuged at 450 g for 5 min (RT), and then re-suspended in 300 µL of PBS for the acquisition of 10,000 events on a cytometer ow (FACS Calibur, Immuno uorometry Systems, Mountain View, CA, USA). Data analysis was performed using BD FACS DIVA 7.0 software (San Jose, CA, USA). Cells were considered to be ECFC when they were positive for CD31, CD144, CD146, and KDR markers, negative for CD45 and CD133, and showed decreased CD34 expression.
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