Splenocytes were prepared as described before. Afterwards, cells were stained for CD3e PerCP-eFluor 710 (clone 17A2, Thermo Fisher Scientific), CD19 FITC (clone 1D3, BD Biosciences), CD11c APC (HL3, BD Biosciences), CD11b PE-Cy7 (clone M1/70, Thermo Fisher Scientific) and HLA-DR BV605 (clone G46-6, BD Biosciences) and subsequently sorted using a BD FACS Aria III (BD Biosciences). Post sort analysis using FlowJo software (Version 10.8.1; BD Biosciences) showed a purity of >98% for splenic CD19+ B cells.
Hla dr bv605 clone g46 6
Manufactured by BD
The HLA-DR BV605 (clone G46-6) is a flow cytometry reagent used for the detection and quantification of HLA-DR molecules on the surface of cells. It is a fluorochrome-conjugated monoclonal antibody that binds to the HLA-DR antigen, which is a major histocompatibility complex (MHC) class II cell surface receptor.
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Lab products found in correlation
3 protocols using hla dr bv605 clone g46 6
1
Splenic B Cell Isolation Protocol
2
Isolation of Splenic Dendritic Cells
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Splenocytes were prepared as described before. Dendritic cells were enriched by a pre-isolation step using NKp46, CD3e and CD19 MACS beads negative selection (all Miltenyi Biotech, Germany). Subsequently, cells were stained for CD3e PerCP-eFluor 710 (clone 17A2, Thermo Fisher Scientific), CD19 FITC (clone 1D3, BD Biosciences), CD11c APC (HL3, BD Biosciences), CD11b PE-Cy7 (clone M1/70, Thermo Fisher Scientific) and HLA-DR BV605 (clone G46-6, BD Biosciences) and sorted using a BD FACS Aria III (BD Biosciences). Nonspecific binding through Fc gamma receptors was blocked by a mixture of anti-CD16 and anti-CD32 antibodies (Fc Block; BD Biosciences). Post sort analysis using FlowJo software (Version 10.8.1; BD Biosciences) showed an average purity of 94.8% for splenic CD11c high MHC class II+ dendritic cells.
3
Enrichment and Characterization of Splenic Monocytes
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Splenocytes were prepared as described before. Monocytes/macrophages were enriched by a pre-isolation step using NKp46, CD3e and CD19 MACS beads negative selection (all Miltenyi Biotech, Germany). Subsequently, cells were stained for CD3e PerCP-eFluor 710 (clone 17A2, Thermo Fisher Scientific), CD19 FITC (clone 1D3, BD Biosciences), CD11c APC (HL3, BD Biosciences), CD11b PE-Cy7 (clone M1/70, Thermo Fisher Scientific) and HLA-DR BV605 (clone G46-6, BD Biosciences) and sorted using a BD FACS Aria III (BD Biosciences). Nonspecific binding through Fc gamma receptors was blocked by a mixture of anti-CD16 and anti-CD32 antibodies (Fc Block; BD Biosciences). Post sort analysis using FlowJo software (Version 10.8.1; BD Biosciences) showed an average purity of 94.4% for splenic CD11b+ monocytes/macrophages.
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