OMIP-023
10-Color, 13 antibody panel for in-depth phenotyping of human peripheral blood leukocytes
Cytometry PART A, Volume 85, Issue 9, 781-784 (2014)
Jozsef Bocsi, Susanne Meizer, Ingo Dahnert, Attila Tarnok
Description: This panel was developed and optimized to determine the phenotype and activation of 15 different leukocyte subtypes in one run. Leukocytes are identified by expression of CD45 (leu-1) pan leukocyte antigen. Neutrophil (CD16), monocyte (CD14), T- (CD3), B-lymphocyte (CD19), and NK-cell (CD16 and CD56) markers are employed. Special gating strategy is used for subtyping of granulocytes (e.g., eosinophils, neutrophils, basophils). For further T-cell phenotyping, CD4/CD8 markers are used for differentiation of four T-cell subtypes, CD25/CD127 for regulatory T cell identification and CD3/CD16/CD56 for NKT-cells, additionally. Special gating strategies have been developed for B-cell, NK-cell, and monocyte subtyping. For detection and analysis of activation also further activation markers (HLA-DR, CD38, CD25, CD127, and CD69) are analyzed. This panel has been established for analysis of human RBC-lysed EDTA-treated whole blood samples and for cord blood. Since the starting material is fresh EDTA-treated blood, dead cells are probably not an issue. Thus to save one channel for specific staining the vitality staining was not used in the panel.
Species: Human
Tissue: Cord blood, Peripheral blood
Cell Subset: Granulocyte, B cell, T cell, Treg, Nk, Monocyte
Keyword: Activation, Immunoprofiling
Machine: Beckman Coulter Navios
Panel details
| Marker | Species | Clone | Fluorophore | Purpose | Co-expressions groups | Antigen Density |
|---|---|---|---|---|---|---|
| CD56 | Human | PE-Cy7 | NK | NK | Low | |
| CD14 | Human | RMO52 | FITC | Monocyte | monocyte | High |
| CD19 | Human | J3-119 | FITC | B-cell differentiation | B cell | Low |
| CD25 | Human | B1.49.9 | ECD | Treg | Treg, T cell | Low |
| CD38 | Human | LS198.4.3 | PE-Cy5.5 | B-cell differentiation | B cell, T cell, Treg, monocyte | Low |
| CD127 | Human | R34.34 | APC-Alexa 700 | Treg gating | Treg | Low |
| CD4 | Human | SK3 | APC-H7 | CD4 T cell | Treg, T cell | High |
| CD3 | Human | SP34-2 | V500 | T cell | Treg, T cell | High |
| HLA DR | Human | Immu-357 | APC | Monocyte, NK gating | monocyte, T cell | High |
| CD8a | Human | B9.11 | FITC | CD8 T cell | T cell | High |
| CD45 | Human | J.33 | Pacific Blue | Leukocyte | monocyte, T cell, B cell, NK, Treg | High |
| CD16 | Human | 3G8 | PE-Cy7 | Monocyte, NK | monocyte, T cell, NK | Med |
| CD69 | Human | TP1.55.3 | PE | Activation | monocyte, NK, T cell, Treg, B cell | Med |
