Flow cytometric analysis of immune inflammatory cells was performed using cell type-specific monoclonal antibodies on 3-day e-cig aerosol-exposed samples. Briefly, 2.0–4.0 × 105 BAL cells were stained with cell type-specific markers in 1x PBS for 30 min, followed by washing and re-suspension in 0.1 mL of 1x PBS for analysis. Cell-specific markers such as LY6B.2 Alexa fluor488-conjugated antibody for neutrophils (Novus Biologicals Cat# NBP213077AF488), CD8a PE-cy5-conjugated antibody for T-lymphocytes (BD Biosciences 553034), and F4/80 PE-conjugated antibody for macrophages (BioLegend Cat #123109) were used. Total macrophage, neutrophil and CD4a+ and CD8a+ lymphocyte counts in BALF were determined by taking the percentage of each cell type observed and multiplying that by the total cell counts. Flow cytometry data acquisition was performed using a Guava® easyCyte™ flow cytometer (Millipore Sigma) and analyzed using Guava® InCyte™ software.
Cd8a pe cy5 conjugated antibody for t
Manufactured by BD
The CD8a PE-cy5-conjugated antibody is a fluorescently labeled monoclonal antibody that binds to the CD8a surface marker on T cells. This antibody can be used for the identification and analysis of T cell populations in flow cytometry applications.
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2 protocols using cd8a pe cy5 conjugated antibody for t
1
E-Cigarette Aerosol Immunophenotyping
2
Inflammatory Immune Cell Profiling
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Cell-type specific monoclonal antibodies were used for flow cytometric analysis of inflammatory immune cells. Briefly, 2.0 × 105 to 4.0 × 105 cells were stained with 1x PBS using cell-type specific markers for 30 min., then washed and re-suspended in 0.1 ml of 1x PBS for analysis. Cell-specific markers such as LY6B.2 Alexa Fluor 488-conjugated antibody for neutrophils (Novus Biologicals Cat# NBP213077AF488) CD8a PE-cy5-conjugated antibody for T-lymphocytes (BD Biosciences 553034) and F4/80 PE-conjugated antibody for macrophages (BioLegend Cat #123109) were used. Flow cytometry data acquisition was performed on a BD Accuri flow cytometer (BD Accuri C6 software) and analyzed using FlowJo software.
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