CD163+CD14+ and CD163−CD14+ populations were sorted from mononuclear cells derived from MPE using Moflo XDP (Beckman) (n=6). In brief, cell clumps were removed by passing cell suspensions through 40 mm Cell Strainers (BD Biosciences). 1×108 mononuclear cells were stained with 20 μl of anti-human CD163, CD14 and 7-AAD antibodies (Biolegend) respectively. Then, cells were incubated in the dark for 15 min at 4 °C. Cells were resuspended with 1 ml of normal saline for sorting. The purities of sorted CD163+CD14+ and CD163−CD14+ cells were analyzed by FACS.
7 aad antibodies
Manufactured by BioLegend
Sourced in United States
7-AAD (7-Aminoactinomycin D) is a fluorescent DNA-binding dye used in flow cytometry to identify dead or dying cells. It is membrane-impermeant and can only enter cells with compromised cell membranes, such as apoptotic or necrotic cells. 7-AAD emits fluorescence upon binding to DNA, allowing for the detection and quantification of these cell populations.
Automatically generated - may contain errors
Lab products found in correlation
Anti human cd163, by BioLegend (1 mentions)
Moflo xdp, by Beckman Coulter (1 mentions)
Cell strainer, by BD (1 mentions)
Trypan blue solution, by Thermo Fisher Scientific (1 mentions)
Cd15 pe, by BioLegend (1 mentions)
Cd11b apc, by BioLegend (1 mentions)
Cytofix cytoperm fixation permeabilization solution kit, by BD (1 mentions)
2 protocols using 7 aad antibodies
1
Sorting CD163+CD14+ and CD163-CD14+ Cells
2
Cell Growth, Cell Cycle, and Apoptosis Analysis
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Cell growth was analyzed at 72 h by counting total cell numbers using trypan blue solution (Thermo Fisher Scientific, Inc., Waltham, MA, USA). For cell cycle analysis, the cells were fixed using the Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (BD Biosciences, NJ, USA) and were incubated with propidium iodide solution. Apoptotic fractions were analyzed by Annexin V/7-AAD staining using the APC Annexin V Apoptosis Detection Kit I (BioLegend, BioLegend, Inc., San Diego, CA, USA). Cell lineage-specific marker expression was measured by flow cytometry using CD15-PE, CD11b-APC, and 7-AAD antibodies (BioLegend, Inc., San Diego, CA, USA).
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