For cell-surface analysis, cells were harvested by dissociation using 0.05% trypsin/EDTA. A total of 1 × 106 cells were resuspended in 200 μL PBS with 1% FBS, incubated with antibodies at the recommended concentrations at 4 °C for 30 min, and then detected by flow cytometry (BD FACSCanto II, BD Biosciences, USA). Data were analyzed using FlowJo software. The antibodies used in flow cytometry assay were as follows: APC-conjugated CD163 antibody (333610, Biolegend), PE-conjugated CD206 antibody (321106, Biolegend), APC-conjugated CD44 antibody (559942, BD Biosciences), PE-conjugated CD24 antibody (555428, BD Biosciences).
Pe conjugated cd24 antibody
Manufactured by BD
The PE-conjugated CD24 antibody is a laboratory reagent used for the detection and analysis of CD24 expression on cells. CD24 is a cell surface glycoprotein that serves as a marker for certain cell types. The PE (phycoerythrin) fluorescent label allows for the visualization and quantification of CD24-positive cells through flow cytometry or other fluorescence-based techniques.
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Lab products found in correlation
Apc conjugated cd44 antibody, by BD (2 mentions)
Pe conjugated cd206 antibody, by BioLegend (1 mentions)
Facscanto 2, by BD (1 mentions)
Apc conjugated cd133 antibody, by Miltenyi Biotec (1 mentions)
Macs cell separation kit, by Miltenyi Biotec (1 mentions)
Moflo xdp cell sorter, by Beckman Coulter (1 mentions)
Anti pe microbeads, by Miltenyi Biotec (1 mentions)
Apc conjugated cd133, by Miltenyi Biotec (1 mentions)
Moflo cell sorter, by Beckman Coulter (1 mentions)
4 protocols using pe conjugated cd24 antibody
1
Cell Surface Marker Analysis by Flow Cytometry
2
Multicolor Flow Cytometry Analysis
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APC‐conjugated CD133 antibody was purchased from Miltenyi (cat. no. 130‐113‐184). PE‐conjugated CD24 antibody was obtained from BD Biosciences (cat. no. 555428). FITC‐conjugated CD326 antibody was purchased from STEMCELL (cat. no. 60136FI). Flow cytometry was performed according to the manufacturer's instructions.
3
Magnetic Cell Sorting and Flow Cytometry
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For magnetic cell sorting, cells were labeled with PE-conjugated CD24 anti-body (BD PharMingen, San Jose, CA) followed by anti-PE microbeads (Miltenyi Biotec, Germany). MACS was performed according to the manufacturer's instructions of Miltenyi Biotec MACS Cell Separation Kit (Miltenyi Biotech Inc, Bergisch Gladbach, Germany). Cells were repeatedly passing through the cell sorting column for at least three times (in order to reach >95% purity of the target cell population). For Flow cytometry cell sorting,was performed using MoFlo XDP Cell Sorter (Beckman Coulter, Fullerton, CA). Cells were incubated with 1 μg of each antibody for 30 min. Control experiments involved incubation with each antibody for 30 min and no apparent increase in the number of dead cells detected by propidium iodide (PI) staining. Flowjo v7.6.2 software (Treestar Inc, San Carlos, CA) was used for data analysis.
4
Isolation of CAIX+ and CAIX- Subpopulations
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Flow cytometry was carried out as previously described [28 (link)]. Samples were analyzed using a DakoCytomation Cyan machine, or sorted using a MoFlo cell sorter (Beckman Coulter). The bottom 10% of CAIX negative subpopulation and the top 10% of CAIX positive subpopulation were collected for further experiments. The following primary antibodies were used FITC conjugated CAIX (R&D systems), APC conjugated CD133 (Miltenyi Biotec), APC conjugated CD44 antibody (BD Biosciences) and PE conjugated CD24 antibody (BD Biosciences).
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