Cd16 pe

Manufactured by Beckman Coulter

Sourced in United States, United Kingdom, France

The CD16-PE is a flow cytometry reagent that detects the CD16 surface marker on cells. It is a monoclonal antibody conjugated with the fluorescent dye phycoerythrin (PE). This reagent is used for the identification and enumeration of CD16-positive cells in biological samples.

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4 protocols using cd16 pe

Phenotypic analysis of dendritic cells

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Frozen PBMC were thawed in RPMI-50% FCS, counted in Trypan blue, and 1 × 10⁶ cells were stained with a viability dye (Viobility 405/520, Fixable Dye, Miltenyi-Biotech), Lineage antibodies (anti-CD3-BV510 (BD Biosciences), CD19-VioGreen (Miltenyi-Biotech) and CD56-BV510 (BioLegend)), and anti-CD14-BV650 (Biolegend), HLA-DR VioBlue (Miltenyi-Biotech), CD141-BV785 (BioLegend), CD16-PE (Beckman-Coulter), CD1c-BV605 (BD Biosciences), FcERI-PerCP-Vio700 (Miltenyi-Biotech), CD123-APC-Vio770 (Miltenyi-Biotech), CD11c-PE-Vio615 (Miltenyi-Biotech) antibodies (all antibodies are described in Additional file 1: Table S3). Samples were processed on the BD LSRFortessa cytometer (BD Biosciences). pDC and DC cells were identified as described by Mair et al. [49 (link)]. Data were analyzed using FlowJoTM version v10·8 software (BD Life Sciences).

Garcia G., Labrouche-Colomer S., Duvignaud A., Clequin E., Dussiau C., Trégouët D.A., Malvy D., Prevel R., Zouine A., Pellegrin I., Goret J., Mamani-Matsuda M., Dewitte A, & James C. (2024). Impaired balance between neutrophil extracellular trap formation and degradation by DNases in COVID-19 disease. Journal of Translational Medicine, 22, 246.

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Immunofluorescence Staining and FACS Analysis of PBMCs

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PBMCs from 43 AD patients and 41 normal subjects were isolated by FACS to further validate the results of single-cell sequencing. Briefly, all PBMCs were washed, counted and suspended in ice-cold PBS, and cell-surface antigen staining was subsequently performed. Monoclonal antibodies against CD3, CD45, CD56, CD19 (CD45/CD56/CD19/CD3 detection kit, Beckman Coulter, USA) and CD16 (CD16-PE, Beckman Coulter, USA) were used in immunofluorescence staining. The stained cells were analyzed and sorted by BD Influx (BD, USA), and data were analyzed using CytExpert 2.0.

Xiong L.L., Xue L.L., Du R.L., Niu R.Z., Chen L., Chen J., Hu Q., Tan Y.X., Shang H.F., Liu J., Yu C.Y, & Wang T.H. (2021). Single-cell RNA sequencing reveals B cell–related molecular biomarkers for Alzheimer’s disease. Experimental & Molecular Medicine, 53(12), 1888-1901.

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Quantification of HIV viral load and lymphocyte subsets

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HIV viral load was measured using branched-chain DNA assay (Siemens Healthcare, Sudbury, UK) with a lower limit of detection of 50 copies/ml, and quantitation of LSS was undertaken by staining whole blood (EDTA) with monoclonal antibodies [CD45-FITC, CD4-RD1, CD8-ECD, CD3-PC5, CD56-RD1, CD19-ECD, CD16-PE (Beckman Coulter Inc., High Wycombe, UK)] and evaluated using FC500 flow cytometer (Beckman Coulter Inc.) via NHS laboratory services. Serological assays were performed in the WHO reference laboratory for pneumococcal serology at the Institute of Child Health, London, UK. Pneumococcal capsular polysaccharide-specific IgG for all PCV13 strains was measured by ELISA after absorption with cell wall and 22F polysaccharide. Opsonophagocytic assay (OPA) was performed according to methods of Romero-Steiner using the promyelocytic HL60 cell line. Detailed protocols for both methods are available at http://www.vaccine.uab.edu/.

Bamford A., Kelleher P., Lyall H., Haston M., Zancolli M., Goldblatt D, & Kampmann B. (2014). Serological response to 13-valent pneumococcal conjugate vaccine in children and adolescents with perinatally acquired HIV infection. AIDS (London, England), 28(14), 2033-2043.

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Immune Cell Profiling in Vedolizumab Response

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Absolute counts of T cells, B cells, NK cells and monocytes were determined by flow cytometry (BD FacsCanto II cytometer with DIVA software) and Sysmex XS-800i analyzer in responder and non-responder patients at inclusion, before the first infusion of vedolizumab, and at week 22. In the same way, α4β7 expression was analyzed by flow cytometry using a FITC-conjugated vedolizumab antibody in the same cells and in B cell subpopulations i.e. Plasmablasts, Transitional B cells, Switch and non-Switch B cells, Naïve B cells.
Within a maximum of 4h after drawing, EDTA whole blood samples (50μl) were incubated 15min with the following antibodies to analyze T cells, B cells, monocytes and NK cells: CD45-KrOrange, CD16-PE, CD14-PECy5, IgG1-FITC (Beckman Coulter, France) and with CD3-BV421, IgG1-APC, CD56-PE, CD19-PECy7, CD8-APC-H7 (BD Biosciences, France). For B cells subpopulation analysis, the following antibodies were used for staining 100μl of washed blood: CD45-KrOrange, IgG1-FITC (Beckman Coulter) and CD24-PE, CD27 PerCP-Cy5, CD38-APC, IgD-APC-H7, CD19-PECy7 (BD Biosciences). Quantitation of lymphocyte subsets (B cells, T cells, CD4 and CD8 T cells and NK cells) was performed using flow cytometry with BD Trucount™ Tubes (BD Biosciences).

Quénéhervé L., Trang-Poisson C., Fantou A., Flamant M., Durand T., Bouguen G., Bregeon J., Oullier T., Amil M., Dewitte M., Bardot S., Blandin S., Braudeau C., Vibet M.A., Josien R., Neunlist M, & Bourreille A. (2024). Confocal laser endomicroscopy as predictive biomarker of clinical and endoscopic efficacy of vedolizumab in ulcerative colitis: The DETECT study. PLOS ONE, 19(4), e0298313.

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