Surface marker analysis by FACS was carried out as described previously [16 (link)]. In brief, hMSC and ES cell lines (RD-ES, SK-N-MC and EWS-GFP) were harvested, centrifugated and incubated at 4°C for 1 h with fluorochrome conjugated antibodies APC Mouse anti-human CD13 (BD Pharmingen, 557454), APC Mouse anti-human CD44 (BD Pharmingen, 560532), APC Mouse anti-human CD73 (BD Pharmingen, 560847), APC Mouse anti-human CD90 (BD Pharmingen, 559869) and APC Mouse anti-human CD105 (BD Pharmingen, 562408). Negative control cells were stained with APC mouse IgG1, k isotype control, Clone MOPC-21 (BD Pharmingen, 555751). CD99 expression was assayed incubating cells with CD99 primary antibody (Signet antibodies, SIG-3620). FACS data were analyzed using FlowJo software version 7.6 (Tree Star Inc., Ashland, OR, USA)
Apc mouse anti human cd105
Manufactured by BD
Sourced in United States
APC mouse anti-human CD105 is a fluorescently labeled antibody that recognizes the human CD105 (endoglin) cell surface antigen. CD105 is expressed on endothelial cells and is involved in angiogenesis and cell proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Pe mouse anti human cd90, by BD (2 mentions)
Pe cy 7 mouse anti human cd44, by BD (2 mentions)
V450 mouse anti human cd31, by BD (2 mentions)
V500 mouse anti human cd45, by BD (2 mentions)
Percp cy 5.5 mouse anti human cd73, by BD (2 mentions)
V450 mouse anti human cd144, by BD (2 mentions)
Cytofix fixation buffer, by BD (2 mentions)
Stain buffer, by BD (2 mentions)
Percp cy5.5 mouse anti human cd90, by BD (2 mentions)
Pe mouse anti human cd73, by BD (2 mentions)
Fitc mouse anti human cd31, by BD (2 mentions)
Facscanto 2, by BD (2 mentions)
Apc mouse antihuman cd73, by BD (1 mentions)
Apc mouse igg1, by BD (1 mentions)
Clone mopc 21, by BD (1 mentions)
Apc mouse anti human cd13, by BD (1 mentions)
Apc mouse anti human cd44, by BD (1 mentions)
Apc mouse anti human cd90, by BD (1 mentions)
Pe mouse anti human cd18, by BD (1 mentions)
Pe mouse anti human cd45, by BD (1 mentions)
6 protocols using apc mouse anti human cd105
1
Surface Marker Analysis of Mesenchymal and Cancer Cells
2
Phenotypic Characterization of OASCs
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Related cell markers of OASCs were analyzed by flow cytometry. The dissociated cells were incubated with fluorescein isothiocyanate (FITC) mouse anti-human CD29, phycoerythrin (PE) mouse anti-human CD34, PE mouse anti-human CD18, FITC mouse anti-human CD49e, PE mouse anti-human CD166, allophycocyanin (APC) mouse anti-human CD133, PE mouse anti-human CD45, and APC mouse anti-human CD105 (BD Biosciences) respectively at 4 °C for 30 min, washed, and resuspended with PBS. The cells then underwent flow cytometry using the BD FACS Calibur. Analysis was performed using the Flow-Jo program (Treestar, USA).
3
Characterization of FGF-17-Modulated hWJ-MSCs
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Normoxic hWJ-MSCs treated with rFGF-17 and transfected with siFGF-17 at passage 7 or hypoxic hWJ-MSCs treated with rFGF-17 and transfected with siFGF-17 at passage 10 were harvested and washed with 1×PBS (Intron Biotechnology, Seoul, Korea). Normoxic hWJ-MSCs not treated with rFGF-17 and transfected with negative control siRNA or hypoxic hWJ-MSCs not treated with rFGF-17 and transfected with negative control siRNA were used as respective control groups. Cells were fixed with BD Cytofix Fixation Buffer (BD Biosciences, Piscataway, NJ, USA) and stained with V450 mouse anti-human CD31 (1:20), fluorescein isothiocyanate (FITC) mouse anti-human CD34 (1:20), phycoerythrin (PE)-Cy™7 mouse anti-human CD44 (1:20), V500 mouse anti-human CD45 (1:20), PerCP-Cy™5.5 mouse anti-human CD73 (1:20), PE mouse anti-human CD90 (1:20), APC mouse anti-human CD105 (1 : 20), and V450 mouse anti-human CD144 (1:20) (BD Biosciences) antibodies for 30 min at room temperature. Cells were washed twice with Stain Buffer (BD Biosciences) and analyzed with a FACSVerse™ flow cytometer (BD Biosciences) and Flowjo software (Treestar, San Carlos, CA, USA).
4
Characterization of Mesenchymal Stem Cells
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MSCs were immunophenotypically characterised by flow cytometry on a BD FACS Canto II (BD Biosciences). As the negative control we used 106 unstained MSCs. Test samples contained 106 MSCs with PE Mouse Anti-Human CD73, APC Mouse Anti-Human CD105, PerCP-Cy5.5 Mouse Anti-Human CD90, FITC Mouse Anti-Human CD31 and PerCP-Cy7 Mouse Anti-Human CD45 (all BD Biosciences). In addition, FACS analysis was used to determine the effect of NMP on the survival of MSCs during perfusion without a kidney with pre-labelled MSCs. For this purpose, hourly perfusate samples were taken. The MSCs could be identified by their fluorescence emission at two different wavelengths as a result of the PKH26 and Q-tracker 655 pre-labelling.
Flow cytometry was also performed on perfusate samples and enzymatically disrupted kidney biopsies from the experiments with pre-labelled MSCs with a kidney in the perfusion circuit. Of each sample, 1 million counts were obtained and analysed through flow cytometry.
Flow cytometry was also performed on perfusate samples and enzymatically disrupted kidney biopsies from the experiments with pre-labelled MSCs with a kidney in the perfusion circuit. Of each sample, 1 million counts were obtained and analysed through flow cytometry.
5
Characterization of Hypoxic Stem Cells
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Normoxic and hypoxic hUC-MSCs at passage 6 were harvested, fixed with BD Cytofix Fixation Buffer (BD Biosciences, Piscataway, NJ, USA) for 10 min at room temperature and washed with Perm/Wash buffer (BD Biosciences). Cells were stained with V450 mouse anti-human CD31 (1:20), fluorescein isothiocyanate (FITC) mouse anti-human CD34 (1:20), phycoerythrin (PE)-Cy™7 mouse anti-human CD44 (1:20), V500 mouse anti-human CD45 (1:20), PerCP-Cy™5.5 mouse anti-human CD73 (1:20), PE mouse anti-human CD90 (1:20), APC mouse anti-human CD105 (1:20), or V450 mouse anti-human CD144 (1:20) (BD Biosciences) antibodies for 30 minutes at room temperature. Cells were washed with Stain Buffer (BD Biosciences), and analyzed with a Caliber flow cytometer (BD Biosciences) using CellQuest program (BD Biosciences) and Flowjo software (Treestar, San Carlos, CA, USA).
6
Multiplex Flow Cytometry for MSC Characterization
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To confirm that the cells cultured were indeed MSCs, we performed flow cytometric analysis on the cells that were used during the experiments on a BD FACS Canto II (BD Biosciences Nederland BV, Vianen, The Netherlands). MSCs were stained with FITC Mouse Anti-Human CD31, PE Mouse Anti-Human CD73, PerCP-Cy5.5 Mouse Anti-Human CD90, and APC Mouse Anti-Human CD105 (all BD Biosciences Nederland BV). As negative control, unstained MSCs were used.
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