10 μL of each antibody (CD3-FITC, CD8-FITC, CD16-FITC, Immunostep, Salamanca, Spain) (CD4-PE, CD20-PE, CD56-PE, Beckman-Coulter, Marseille, France) were added to 100 μL whole blood in tubes. Isotype controls included the replacement of specific antibodies with isotype mouse immunoglobulins. After vortexing and incubation for 30 min at room temperature in dark, 2 mL lysing solution was added and RBCs were lysed for 10 min at room temperature. Samples were centrifuged (Hettich Zentrifugen, Switzerland) for 5 min at 460 ×g. After pelleting, 2 mL washing solution was added and samples were centrifuged for 5 min. Prior to analysis, leukocytes were resuspended in 1ml PBS containing 1% formaldehyde. Samples were analyzed using Flow Cytometer device (Partec Cube 6) and FCS express Software. Ten thousand events were analyzed per sample. Positive populations were identified using Isotype control.
Cd20 pe
Manufactured by Beckman Coulter
Sourced in Spain
CD20-PE is a fluorescently-labeled monoclonal antibody that binds to the CD20 antigen expressed on the surface of B cells. It is used in flow cytometry applications for the identification and enumeration of B cells in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Cd56 pe, by Beckman Coulter (2 mentions)
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Abx pentra 60 hematology analyzer, by Horiba (1 mentions)
2 protocols using cd20 pe
1
Multiparameter Flow Cytometry Analysis
2
Immune Cell Profiling in Leukapheresis
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The quantity and percentage of various immune cell types in leukapheresis product and elutriation fractions were analyzed by an ABX Pentra 60 hematology analyzer (HORIBA). For other experiments, FACS analysis (Accuri C6 Plus; Beckman Coulter Life Sciences) was used. Monocytes were enumerated by CD45 + /CD14 + and propidium iodideÀnegative population (Beckman Coulter Life Sciences). Dendritic cells were gated by forward scatter and side scatter on the basis of cell size and cellular complexity, and further gated by anti-CD45 and propidium iodide. Phenotypic analyses of dendritic cells were performed using anti-CD209-APC, CD80-PE, CD83-PE, CD86-PE, CD11c-PE, HLA-DR-APC, HLA-ABC-APC and CCR7-APC (Beckman Coulter Life Sciences). To analyze cell type compositions in CUD-002 product, anti-CD45-APC, CD19-PE, CD20-PE, CD3-Percp-Cy5.5, CD56-PE, CD34-PE, lineage cocktail (CD3/CD14/CD16/CD19/CD20/CD56)-FITC, CD13-PE, CD33-PE and S100A9-PE (Beckman Coulter Life Sciences) were used.
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