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OMIP-051

28‐color flow cytometry panel to characterize B cells and myeloid cells

Thomas Liechti, Mario Roederer Cytometry PART A, Volume 95, Issue 2, 105-155 (2019)

PURPOSE: This 28‐color flow cytometry panel focuses on B cells, dendritic cells, and monocytes and was optimized for cryopreserved peripheral blood mononuclear cells (PBMC). In addition to markers enabling the analysis of monocytes (CD14) and definition of subsets within B cells (CD10, CD19, CD20, CD21, CD27, IgD, IgM) and dendritic cells (CD1c, CD11c, CD123, CD141, HLA‐DR), we included functional markers such as chemokine receptors (CXCR3, CXCR5), surface immunoglobulins (IgA, IgD, IgG, IgM), Fc receptors (CD16, CD23, CD32, CD64), inhibitory molecules (CD73, CD85j), the co‐stimulatory molecule CD40, and cytokine receptors (IL‐21R, BAFF‐R, TACI), which enables in‐depth characterization of B cells, dendritic cells, and monocytes.

CELL TYPE: PBMC

MACHINE: BD FACSymphony A5

Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Blue viability'
CD73 Human AD2 BB515 Inhibitory
CD141 Human 1A4 BB630-P2 DC lineage marker
CD123 Human 7G3 BB660-P2 DC lineage marker
CD16 Human 3G8 BB700 Fc receptor
IgD Human IA6-2 BB790-P Differentiation B cells
CD32 Human FUN-2 PE Fc receptor
CD40 Human 5C3 PE-Dazzle 594 Co-stimulatory
CD85j Human GHI/75 PE-Cy5 inhibitory
CD11c Human Bu15 PE-Cy5.5 DC lineage marker
CD183 Human G025H7 PE-Cy7 Trafficking
IgA Human polyclonal APC Differentiation B cells
CD27 Human M-T271 APC-R700 Differentiation B cells
CD19 Human SJ25C1 APC-H7 Lineage B cell
CD1c Human BUV395 DC lineage marker
CD21 Human B-ly4 BUV496 Differentiation B cells
CD267 Human 1A1-K21-M22 BUV563 Cytokine receptor
HLA-DR Human G46-6 BUV661 DC lineage marker
IgG Human BUV737 Differentiation B cells
CD20 Human 2H7 BUV805 Lineage B cell
IL-21 R Human 2G1-K12 Brilliant Violet 421 Cytokine receptor
CD14 Human Brilliant Violet 510 Lineage monocyte
IgM Human MHM-88 Brilliant Violet 570 Differentiation B cells
CD268 Human 11C1 Brilliant Violet 605 Cytokine receptor
CD10 Human HI10a Brilliant Violet 650 Differentiation B cells
CD23 Human M-L233 Brilliant Violet 711 Differentiation B cells
CD185 Human RF8B2 Brilliant Violet 750 Trafficking
CD64 Human 10.1 Brilliant Violet 786 Fc receptor