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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.

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2 protocols using 7 aad antibodies

1

Sorting CD163+CD14+ and CD163-CD14+ Cells

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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.
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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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