Following selection with puromycin, GMan-derived cells (1.0×105) were suspended in phosphate-buffered saline supplemented with 0.5% FBS and 2 mM EDTA. The cells were incubated with fluorescein isothiocyanate (FITC)-conjugated anti-mouse Sca-1 (1:10; 130-102-297), anti-mouse CD44 (1:10; 130-102-511), or anti-mouse CD90 (1:10; 130-102-452) antibodies for 1 h at 4°C in the dark. All FITC-conjugated antibodies were purchased from Miltenyi Biotec GmbH (Bergisch Gladbach, Germany). Image acquisition was performed with an EPICS XL ADC system (Beckman Coulter, Inc., Brea, CA, USA).
Epics xl adc system
Manufactured by Beckman Coulter
Sourced in United States
The EPICS XL ADC system is a flow cytometry instrument designed for cell analysis and sorting. It is equipped with a multi-color detection system and digital signal processing technology for rapid data acquisition and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Cellquest software, by BD (1 mentions)
Epics xl flow cytometer, by Beckman Coulter (1 mentions)
Annexin 5 fitc pi kit, by BD (1 mentions)
Fitc conjugated anti stro 1, by BioLegend (1 mentions)
Fitc conjugated anti cd105, by BioLegend (1 mentions)
Fitc conjugated anti cd271, by BioLegend (1 mentions)
Fitc conjugated anti cd90, by BioLegend (1 mentions)
6 protocols using epics xl adc system
1
Immunophenotyping of GMan-derived Cells
2
Quantifying Cellular Apoptosis by Flow Cytometry
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To quantify cellular apoptosis in the early and late stages, cells were seeded in a 6-well plate, and after treatment, they were harvested and processed for the apoptosis assay using an Annexin V-FITC/PI kit (BD, San Jose, CA, United States) following the indicated protocol specified by the manufacturer. They were examined using an EPICS XL flow cytometer and EPICS XL ADC system (Beckman Coulter, Porterville, CA, United States). Data were analyzed using Cell Quest software (BD, San Jose, CA, United States).
3
Multiparametric Flow Cytometry Analysis
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Cells were suspended in PBS containing 0.5% FBS and 2 mM EDTA at (1.0 × 105) and incubated with phycoerythrin (PE)-conjugated anti-BST1 (clone: eBioSY11B5, eBioscience, 12-1579-4), fluorescent isothiocyanate (FITC)-conjugated anti-CD271 (clone: ME204, BioLegend, 345103), FITC-conjugated anti-CD90 (clone: 5E10, BioLegend, 328107), PE-conjugated anti-CD106 (clone: STA, BioLegend, 305805), FITC-conjugated anti-Stro-1 (BioLegend, 340105), PE-conjugated anti-MSCA-1 (clone: W8B2, BioLegend, 327305), FITC-conjugated anti-CD105 (clone: 43A3, BioLegend, 323203), FITC-conjugated anti-CD73 (clone: AD2, BioLegend, 344015), or PE-conjugated anti-CD146 (clone: SHM-57, BioLegend, 342003) antibody for 1 h at 4°C in the dark. Acquisition was performed with an EPICS XL ADC System (Beckman Coulter).
4
Phenotypic Analysis of SG2 Cells
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SG2 cells directly or indirectly cocultured with SCDC2 cells were suspended in ice-cold phosphate-buffered saline (PBS) containing 0.5% FBS and 2 mM ethylenediaminetetraacetic acid (EDTA). The cells were incubated with phycoerythrin- (PE-) conjugated anti-mouse SCA-1, CD44, and CD90 antibodies (Miltenyi Biotec) for 1 h at 4°C in the dark. Acquisition was performed with an EPICS XL ADC System (Beckman Coulter, Brea, CA, USA). Measurement of fluorescence intensity relative to PE in SG2 cells was performed after gating according to the fluorescence of GFP.
5
Characterization of Bone Marrow-Derived Cell Lines
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A total of 1.0×105 bone marrow-derived cell lines (SG-2, -3, -5 and -6) were suspended in PBS containing 0.5 FBS and 2 mM EDTA and incubated with phycoerythrin-conjugated monoclonal anti-mouse Sca-1 (1:10; cat. no. 130-093-224), monoclonal anti-mouse CD44 (1:10; cat. no. 130-096-838), monoclonal anti-mouse CD11b (1:10; cat. no. 130-091-240) or monoclonal anti-mouse CD45 (1:10; cat. no. 130-091-610) antibody for 1 h at 4°C in the dark. All antibodies were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany). Acquisition was performed using an EPICS XL ADC system (Beckman Coulter, Brea, CA, USA).
6
VEGFR3 Expression in Cell Lines
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Almost confluent SG-2, SG-3 and SG-5 cells (1.0×105) were suspended in ice-cold phosphate-buffered saline (PBS) containing 0.5% FBS and 2 mM EDTA. The cells were incubated with phycoerythrin (PE)-conjugated anti-mouse VEGFR3 (CD310) antibody (1:10, clone AFL4, #130-102-216; Miltenyi Biotec, Bergisch Gladbach, Germany) for 1 h at 4°C in the dark. Acquisition was performed with an EPICS XL ADC system (Beckman Coulter, Inc., Brea, CA, USA).
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