Cell surface marker expression was evaluated using the following primary antibodies: the stage-specific embryonic antigen 4-PE (SSEA4-PE; BioLegend, Dedham, MA, USA), CD73-APC (Thermo Fisher Scientific), CD90-APC-Vio770 (Miltenyi Biotec, San Diego, CA, USA), CD105-PerCP-Vio700 (Miltenyi Biotec), and CD146-PE (Thermo Fisher Scientific). Expanded cells (2 × 105 each group) were first blocked through a 30 min incubation in cold phosphate-buffered saline (PBS) containing 0.1% ChromPure Human IgG whole molecule (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) followed by incubation with primary antibodies at 4 °C for 30 min. Fluorescence was evaluated by a FACS Calibur (BD Biosciences) using the FCS Express software package (De Novo Software).
Ssea4 pe
Manufactured by BioLegend
Sourced in United States
SSEA4-PE is a fluorescent-conjugated antibody used for the detection of the SSEA4 antigen. SSEA4 is a carbohydrate epitope expressed on the surface of embryonic stem cells and other pluripotent cells. The PE (Phycoerythrin) fluorescent label allows for the visualization and quantification of SSEA4-positive cells using flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur, by BD (3 mentions)
Fcs express software, by De Novo Software (2 mentions)
Cd73 apc, by Thermo Fisher Scientific (2 mentions)
Cd90 apc vio770, by Miltenyi Biotec (2 mentions)
Cd105 percp vio700, by Miltenyi Biotec (2 mentions)
Cd146 pe, by Thermo Fisher Scientific (2 mentions)
Chrompure human igg whole molecule, by Jackson ImmunoResearch (2 mentions)
Facsaria 1, by BD (1 mentions)
Cd34 pe, by BioLegend (1 mentions)
Anti human cd31 fitc, by Thermo Fisher Scientific (1 mentions)
Vegfr2 kdr pe, by R&D Systems (1 mentions)
Cd144 fitc, by Abcam (1 mentions)
Cd90 pe, by BioLegend (1 mentions)
Cellquest software, by BD (1 mentions)
Cd14 fitc, by Thermo Fisher Scientific (1 mentions)
Cd29 pe, by BioLegend (1 mentions)
Cd45 fitc, by BioLegend (1 mentions)
Cd34 pe, by BD (1 mentions)
Dapi 4 6 diamidino 2 phenylindole, by Thermo Fisher Scientific (1 mentions)
Cd45 bv421, by BD (1 mentions)
6 protocols using ssea4 pe
1
Characterizing Mesenchymal Stem Cell Markers
2
Phenotypic Characterization of Endothelial Progenitor Cells
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The EPC single-cell suspension was generated into the concentration of 1×107 cells/ml. The cells were then incubated respectively with anti-human CD31-FITC (eBioscience, San Diego, CA, USA), vascular endothelial growth factor receptor (VEGFR2)/KDR-PE (R&D Systems, Minneapolis, MN, USA), CD144-FITC (Abcam, Cambridge, UK), CD34-PE, CD45-FITC (both from Biolegend, San Diego, CA, USA), CD14-FITC (eBioscience), CD29-PE, CD90-PE and SSEA4-PE (all from Biolegend). Briefly, 100 µl cell suspension was incubated with 5 µl antibody solution at 4°C for 30 min in the dark. After washing twice with phosphate buffer saline (PBS), cells were resuspended in 400 µl PBS and analyzed with a FACSAria I (Becton-Dickinson, San Jose, CA, USA) and Becton-Dickinson CellQuest software.
3
Flow Cytometric Analysis of Stem Cell Markers
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hESCs or HCT116 cells were collected and washed in cold PBS, and then cells were incubated with primary antibodies against CD133-APC (Miltenyi Biotec, 130-098-129) or SSEA-4-PE (BioLegend, 330,405) and incubated for 30 min on ice. Samples were washed three times with PBS and analysis was performed using a flow cytometer (BD FACS Calibur).
4
Multiparametric Flow Cytometry for Lineage Tracing
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Cellular composition of the liquid cultures and in vivo tissues were analyzed using flow cytometry. The following antibodies and recombinant proteins were used: Sca1-Pacific blue (PB), CD34-Phycoerythrin (PE), CD48-PE, CD150-PE.Cy7, Ter119 Allophycocyanin (APC)(all Biolegend), CD117-APC, CD16/32-APC.Cy7, CD11b-PE, Gr1-APC, CD3-PE.Cy7, CD71 PE (all BD Biosciences), CD19 Alexa Fluor 700 (AF700, Life) and CD117 PerCpCy5.5 (SONY). Lineage positive cells were detected using the murine lineage detection kit (BD Biosciences) subsequently stained with streptavidin-pacific orange (PO, Fisher Scientific).
SSEA4-PE (Biolegend) and Tra-1-60 Alexa Fluor 647 (AF647, BD Biosciences) were used for regular pluripotency screenings of the iPSCs. Hematopoietic induction of iPSC was assessed using CD34-PE and CD45-BV421 antibodies (both BD Biosciences). Dead cells were excluded by using 7-aminoactinomycin-D (7AAD, Life) or 4’,6-Diamidino-2-Phenylindole (DAPI, Fisher Scientific). Flow cytometry was performed using a BD LSRII flow cytometer (BD Biosciences). For subsequent cell sorting a FACSAria (BD Biosciences) was used. Analysis of FACS data was performed using FlowJo (TreeStar).
SSEA4-PE (Biolegend) and Tra-1-60 Alexa Fluor 647 (AF647, BD Biosciences) were used for regular pluripotency screenings of the iPSCs. Hematopoietic induction of iPSC was assessed using CD34-PE and CD45-BV421 antibodies (both BD Biosciences). Dead cells were excluded by using 7-aminoactinomycin-D (7AAD, Life) or 4’,6-Diamidino-2-Phenylindole (DAPI, Fisher Scientific). Flow cytometry was performed using a BD LSRII flow cytometer (BD Biosciences). For subsequent cell sorting a FACSAria (BD Biosciences) was used. Analysis of FACS data was performed using FlowJo (TreeStar).
5
Endothelial Marker Expression Analysis
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Flow cytometry was performed to analyze the percentage of endothelial markers expression in different cell samples. Cells were harvested, washed with ice-cold FACS buffer (PBS supplemented with 2% FBS and 5 mM EDTA) and incubated with conjugated antibody for 30 minutes on ice. Compensation was performed with CompBeads (BD). Specific fluorochrome-conjugated antibodies were applied to stain the cell surface: CD34-PerCP (343519, Biolegend); CD34-FITC (343504, Biolegend); CD31-PE (303106, Biolegend); CD309(VEGFR2/KDR)-PE (359903, Biolegend); CD117(C-KIT)-PE (313203, Biolegend); CD140a(PEGFRα)-PE (323505, Biolegend); SSEA4-PE (330405, Biolegend). BD Accuri C6 Cytometer was used to perform flow cytometry. Data was collected and analyzed with FlowJo V10. For double staining analysis, fluorescence-minus-one (FMO) analyses were performed on Control or PR-SMCs to set the threshold for CD34 and CD31/KDR/C-KIT/PDGFRα positivity.
6
Characterizing Mesenchymal Stem Cell Markers
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Cell surface marker expression was evaluated using the following primary antibodies: the stage-specific embryonic antigen 4-PE (SSEA4-PE; BioLegend, Dedham, MA, USA), CD73-APC (Thermo Fisher Scientific), CD90-APC-Vio770 (Miltenyi Biotec, San Diego, CA, USA), CD105-PerCP-Vio700 (Miltenyi Biotec), and CD146-PE (Thermo Fisher Scientific). Expanded cells (2 × 105 each group) were first blocked through a 30 min incubation in cold phosphate-buffered saline (PBS) containing 0.1% ChromPure Human IgG whole molecule (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) followed by incubation with primary antibodies at 4 °C for 30 min. Fluorescence was evaluated by a FACS Calibur (BD Biosciences) using the FCS Express software package (De Novo Software).
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