BMMCs and BMA were analyzed for the expression of cell-surface antigens with direct three-color analysis using fluorescein isothiocyanate (FITC)-conjugated, phycoerythrin (PE)-conjugated and peridinin chlorophyll protein complex (PerCP)-conjugated monoclonal antibodies as previously reported [33 (link), 34 (link)]. The following antibodies were used for analysis: immunoglobulin G1 control PE (eBioscience, San Diego, CA, USA), immunoglobulin G1 control PerCP (Becton Dickinson, San Jose, CA, USA), immunoglobulin G1 control FITC (BD Bioscience, Franklin Lakes, NJ, USA), anti-CD34-FITC (BD Bioscience), anti-CD34-PerCP (Becton Dickinson), anti-KDR-PE (R&D Systems, Minneapolis, MN, USA), anti CD45-PerCP (BD Bioscience), and anti-CD133–PE (Miltenyi Biotec, Bergisch Gladbach, Germany). The quantification of total mononuclear cells in BMMCs and BMA was determined by Turk’s solution. For FACS analysis, 5 × 105 events were acquired and scored with a FACSCalibur analyzer (Becton Dickinson). Data were processed using the Macintosh CELLQuest software program (Becton Dickinson).
Anti kdr pe
Manufactured by R&D Systems
Sourced in United States
Anti-KDR-PE is a laboratory reagent that binds to the KDR (Kinase Insert Domain Receptor) protein. It is conjugated with the fluorescent dye Phycoerythrin (PE) for detection purposes. The primary function of this product is to enable the identification and analysis of KDR-expressing cells in research applications.
Automatically generated - may contain errors
3 protocols using anti kdr pe
1
Multicolor Flow Cytometric Analysis of Bone Marrow
2
Multilineage Cell Surface Antigen Analysis
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BMMCs and BMA were analyzed for the expression of cell surface antigens using direct three-color analysis with fluorescein isothiocyanate (FITC)-conjugated, phycoerythrin (PE)-conjugated, and peridinin chlorophyll protein (PerCP)-conjugated monoclonal antibody complexes as previously reported (Fadini et al., 2007 (link); Nevskaya et al., 2008 (link)). The following antibodies were used for analysis: PE-conjugated immunoglobulin G1 control (eBioscience, San Diego, CA, United States), PerCP-conjugated immunoglobulin G1 control (Becton Dickinson, San Jose, CA, United States), FITC-conjugated immunoglobulin G1 control (BD Bioscience, Franklin Lakes, NJ, United States), anti-CD34-FITC (BD Bioscience), anti-CD34-PerCP (Becton Dickinson), anti-KDR-PE (R & D Systems, Minneapolis, MN, United States), anti-CD45-PerCP (BD Bioscience), and anti-CD133-PE (Miltenyi Biotec, Bergisch Gladbach, Germany). The quantification of total mononuclear cells in CMMOs and BMA was determined by Turk’s solution. For FACS analysis, 5 × 105 events were acquired and analyzed with a FACSCalibur analyzer (Becton Dickinson). Data were processed using the Macintosh CELLQuest software program (Becton Dickinson).
3
Quantification of Endothelial Progenitor Cells
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Hematopoietic stem cells (CD34 and CD133) and endothelial cells (KDR) are the surface markers of EPCs, and the determination of these three markers represents the number of EPCs. Flow cytometry was used to determine the expression of CD34, CD133, and KDR in mononuclear cells (MNCs). A volume of 10 mL fresh peripheral venous blood treated with heparin (20 u/mL) was used to obtain MNCs via a Ficoll-Hypaque Solution (Tianjin Hanyang Biologicals Technology Co., Ltd, China). The MNCs were then re-suspended in a M199 medium (Hyclone, USA) and diluted in a FACS buffer for the measurement of CD34, CD133, and KDR via incubation with RESEARCH anti-CD34-PC-5 (Becton Dickinson, USA), anti-CD133-FITC (Beijing Bo Orson Biological Technology Co., Ltd., China), and anti-KDR-PE (R&D Systems, USA) in the dark. Cells were washed twice with the FACS buffer and fixed with 2% paraformaldehyde. Analysis was performed via the assessment of 100,000 events for each separate examination. The expression of CD34, CD133, and KDR was reported as a percentage of the total events.
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