Expression of stem cell surface markers in DPCs was determined by fluorescence-activated cell sorting (FACS) analysis. The cells were detached using trypsin in 0.25% elhylene diamine tetraacetic acid (EDTA). After neutralization, single-cell suspensions were washed with phosphate-buffered saline supplemented with 2% FBS and 0.01% NaN3 (FACS buffer). Quantities of 1 × 105 cells were incubated with the conjugated antibody for 20 min on ice in the dark. After washing, fluorescence intensity was measured on FACS Aria II cell sorter (BD Biosciences, San Jose, CA, USA). The following antibodies were used: Phycoerythrin (PE)-CD271 (Miltenyi Biotec, Auburn, CA, USA), Fluorescein Isothiocyanate (FITC)-CD90 (Biolegend, San Diego, CA, USA), allophycocyanin (APC)-CD106 (Biolegend, San Diego, CA, USA) or dual color combinations APC-STRO-1/PE-CD146 (Both from: Biolegend, San Diego, CA, USA), and PE-CD51 (Biolegend, San Diego, CA, USA)/APC-CD140α (BD Biosciences, San Jose, CA, USA). PE-IgG was used as a negative control.
Cd51 pe
Manufactured by BioLegend
Sourced in United States
CD51-PE is a fluorescently-labeled monoclonal antibody that recognizes the CD51 antigen. CD51 is a subunit of the vitronectin receptor, which is involved in cell adhesion and migration. The PE (Phycoerythrin) fluorescent label allows for the detection and analysis of CD51-expressing cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Facsaria 2 cell sorter, by BD (2 mentions)
Phycoerythrin pe cd271, by Miltenyi Biotec (2 mentions)
Fluorescein isothiocyanate fitc cd90, by BioLegend (2 mentions)
Allophycocyanin apc cd106, by BioLegend (2 mentions)
Apc stro 1 pe cd146, by BioLegend (2 mentions)
Apc cd140α, by BD (2 mentions)
Collagenase d, by Merck Group (1 mentions)
Cd31 percp cy5, by BioLegend (1 mentions)
Cd45 apc cy7, by BioLegend (1 mentions)
Facsaria 2, by BD (1 mentions)
Cd31 fitc, by BioLegend (1 mentions)
Sca 1 pe cy7, by BioLegend (1 mentions)
Cd45 percp cy5, by BioLegend (1 mentions)
Lin apc, by BD (1 mentions)
5 protocols using cd51 pe
1
Flow Cytometry Analysis of Stem Cell Markers
2
Stem Cell Surface Marker Expression Analysis
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Expression of stem cell surface markers in PDLCs was determined by fluorescent activated cell sorting (FACS) analysis. The cells were detached using trypsin in 0.25% ethylenediaminetetraacetic acid (EDTA). After neutralization, single-cell suspensions were washed with phosphate-buffered saline (PBS) supplemented with 2% FBS and 0.01% NaN3 (FACS buffer). Quantities of 1 × 106 cells were incubated with direct conjugated antibodies for 20 min on ice in the dark. After washing, fluorescence intensity was measured on FACS Aria II cell sorter (BD Biosciences, San Jose, CA, USA). The following anti-human antibodies were used: phycoerythrin (PE)-CD271 (Miltenyi Biotec, Auburn, CA, USA), fluorescein isothiocyanate (FITC)-CD90 (BioLegend, San Diego, CA, USA), allophycocyanin (APC)-CD106 (BioLegend, San Diego, CA, USA) or dual color combinations APC-STRO-1/PE-CD146 (Both from: BioLegend, San Diego, CA, USA), and PE-CD51 (BioLegend, San Diego, CA, USA)/APC-CD140α (BD Biosciences, San Jose, CA, USA). PE-IgG was used as a negative control.
3
Isolation and analysis of osteoblasts and lymphocytes
4
Isolation and Characterization of Murine Mesenchymal Stem Cells
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Mouse tibiae were flushed using a 21 G needle to collect BM cells into digestion buffer containing 2 mg·mL−1 collagenase IV/ 3 mg·mL−1 dispase in 1X PBS. Multiple cycles of digestion with agitation were performed at 37 °C (7 min each) to obtain single cell suspension. Single cell suspensions were transferred into DMEM medium containing 10% calf serum and maintained at 4 °C until the rest of the tissue was completely digested. Red blood cells were lysed in 1:1 NH4Cl hypotonic solution. Cells were filtered into clean 50 mL conical tubes, centrifuged at 1 100 r·min−1, 4 °C, for 10 min, resuspended in staining buffer (2 mmol·L−1 EDTA, 0.5% BSA in 1X PBS), and transferred to FACS tubes for staining. The following cell-surface markers were used: For exclusion hematopoietic cells: TER119-AF700 (Biolegend, Clone:TER119, 1:100) and CD45-APC-Cy7 (Biolegend, Clone:30-F11, 1:200). For identifying MSCs: CD140a-Biostin (Biolegend, Clone:APA5, 1:200) and CD51-PE (Biolegend, clone:RMV-7, 1:200). MSCs (CD45-Ter119-CD31-PDGFRα+ CD51+) were used for colony and gene expression assays.
5
Immunophenotyping of Bone Marrow Cells
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A total of 1 × 107 BM cells collected as described above were resuspended in 100 µl of PBS containing specific anti-mouse antibodies (Lin/APC-BD Biosciences, CD45/PerCP-Cy5.5-Biolegend, CD31/FITC-Biolegend, Sca-1/PE-Cy7-Biolegend and CD51/PE-Biolegend) and incubated for 30 min at room temperature in the dark. Frequency of each cell subset was determined using a FacsAria II (BD Biosciences) with the following antibodies: Multipotent Stromal Cells—MSCs (CD45-/Lin-/CD31-/Sca1+/CD51+); Osteoblastic Lineage Cells—OBCs (CD45-/Lin-/CD31-/Sca1-/CD51+); Arteriolar Endothelial Cells—AEC (CD45-/Lin-/CD31+/Sca1+) and Sinusoidal Endothelial Cells—SEC (CD45-/Lin-/CD31+/Sca1-). Analysis was performed with FlowJo software (TreeStar Inc.).
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