The antibodies used were anti-HLA-DR-FITC, mouse-IgG2b-FITC (both from BD Biosciences, San Jose, CA, USA); anti-CD83-PE, mouse-IgG1-PE (both from BioLegend, San Diego, CA, USA); anti-CD14-PerCP, mouse-IgG1k-PerCP, anti-CD11-APC, mouse-IgG1k-APC, biotinylated antihuman DEC-205 monoclonal antibody, biotinylated mouse IgG2bk (all from eBiosciences, San Diego, CA, USA); antihuman interferon (IFN)-γ enzyme-linked immunosorbent assay kit (BD Pharmingen, San Diego, CA, USA).
Anti cd14 percp
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Anti-CD14-PerCP is a monoclonal antibody conjugated with PerCP (Peridinin Chlorophyll Protein) that binds to the CD14 cell surface marker. CD14 is a glycosylphosphatidylinositol (GPI)-anchored protein expressed on the surface of monocytes, macrophages, and neutrophils. This antibody can be used for the identification and enumeration of cells expressing CD14 through flow cytometry analysis.
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Lab products found in correlation
Anti cd45 fitc, by Thermo Fisher Scientific (2 mentions)
Anti cd105 pe, by Thermo Fisher Scientific (2 mentions)
Anti cd19 apc, by Thermo Fisher Scientific (2 mentions)
Calibur, by BD (2 mentions)
Anti cd90 pe, by Thermo Fisher Scientific (2 mentions)
Pe mouse igg1, by BioLegend (1 mentions)
Anti hla dr fitc, by BD (1 mentions)
Anti cd83 pe, by BioLegend (1 mentions)
Facs flow cytometer, by BD (1 mentions)
Anti cd34 fitc, by Thermo Fisher Scientific (1 mentions)
Anti cd44 apc cy7, by Thermo Fisher Scientific (1 mentions)
Anti cd31 pe cy7, by Thermo Fisher Scientific (1 mentions)
Anti cd105 apc, by Thermo Fisher Scientific (1 mentions)
Anti cd73 apc cy7, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc, by BD (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Annexin 5 apc 7 aad apoptosis detection kit, by BestBio (1 mentions)
4 protocols using anti cd14 percp
1
Multicolor Flow Cytometry for Immune Profiling
2
Phenotypic Characterization of gb-MSCs
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Flow cytometry analysis was performed using fluorochrome-conjugated antibodies. Briefly, subconfluent gb-MSCs were cultured in flasks with serum-free medium (0% DMEM), standard medium containing 10% FBS (10% DMEM), serum-free glioblastoma-conditioned medium (0% gb-CM) and standard glioblastoma-conditioned medium(s-gb-CM) for 72 h. The cells were trypsinized and collected in PBS. After centrifugation, the resuspended cells were stained with fluorochrome-conjugated antibodies, including anti-CD31-PE/Cy7, anti-CD34-FITC, anti-CD73-APC/Cy7, anti-CD90-PE/Cy7, anti-CD14-percp, anti-CD105-APC and anti-CD44-APC/Cy7 (all from ebioscience. USA) as well as anti-PDGFR-β-PE (R&D, USA) in the dark at 4°C for 30 min. Then, the cells were centrifuged, resuspended in PBS and analyzed using a FACS flow cytometer (BD Biosciences). The data were collected and analyzed using FlowJo (TreeStar, Ashland, OR) software.
3
Flow Cytometric Characterization of SHED Cell Surface Markers and Apoptosis Analysis
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The surface markers of SHED cells were examined using a flow cytometer (Calibur, BD Biosciences). 1 × 106 SHED cells were incubated with following fluorochrome-conjugated antibodies: anti-CD44-APC (Cat. MA1-10,226, Invitrogen), anti-CD105-PE (Cat. 12–1057-42, Invitrogen), anti-CD90-PE (Cat. 12–0909-42, Invitrogen), anti-CD45-FITC (Cat. 14–9457-95, Invitrogen), anti-CD19-APC (Cat. 17–0199-42, Invitrogen) and anti-CD14-PerCP (Cat. 450,149–42, Invitrogen) for 1 h at 4℃ in the dark and were then analyzed after washing in PBS.
The percentage of cell apoptosis was determined by FACS. In brief, 3 × 104 HUVECs per well were seeded into six-well plates. Following incubation with SHED-Exos (30 μg/ml) or PBS (control) for 48 h, cells in the supernatant were collected, centrifuged and incubated with 500 μl binding buffer containing 10 μl PI and 5 μl Annexin V-FITC (Cat. 556,419, BD Biosciences) for 10 min at room temperature in the dark and were then analyzed by FACS.
The percentage of cell apoptosis was determined by FACS. In brief, 3 × 104 HUVECs per well were seeded into six-well plates. Following incubation with SHED-Exos (30 μg/ml) or PBS (control) for 48 h, cells in the supernatant were collected, centrifuged and incubated with 500 μl binding buffer containing 10 μl PI and 5 μl Annexin V-FITC (Cat. 556,419, BD Biosciences) for 10 min at room temperature in the dark and were then analyzed by FACS.
4
Flow Cytometric Characterization of PDLSCs
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The surface markers of PDLSCs were examined using a flow cytometer (Calibur, BD Biosciences). 1 × 106 PDLSCs were incubated with following luorochrome-conjugated antibodies: anti-CD44-APC (Cat. MA1-10,226, Invitrogen), anti-CD105-PE (Cat. 12–1057–42, Invitrogen), anti-CD90-PE (Cat. 12–0909–42, Invitrogen), anti-CD45-FITC (Cat. 14–9457–95, Invitrogen), anti-CD19-APC (Cat. 17–0199–42, Invitrogen) and anti-CD14-PerCP (Cat. 450,149–42, Invitrogen) for 1 h at 4°C in the dark and were then analyzed after washing in PBS.
The percentage of cell apoptosis was determined by FACS. Apoptotic PDLSCs were detected according to the instructions of the AnnexinV-APC/7-AAD apoptosis detection kit (BestBio, Shanghan, China). Briefly, PDLSCs were trypsinized and the resuspended. PDLSCs were washed with PBS and stained with annexin V-APC and 7-AAD. The cells were then analyzed by flow cytometry (CytoFLEX; Beckman Coulter, Brea, CA, United States).
The percentage of cell apoptosis was determined by FACS. Apoptotic PDLSCs were detected according to the instructions of the AnnexinV-APC/7-AAD apoptosis detection kit (BestBio, Shanghan, China). Briefly, PDLSCs were trypsinized and the resuspended. PDLSCs were washed with PBS and stained with annexin V-APC and 7-AAD. The cells were then analyzed by flow cytometry (CytoFLEX; Beckman Coulter, Brea, CA, United States).
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