Spleen single-cell suspensions were collected from the mice, filtered with a cell strainer, distributed into 2 sets and were suspended in 50 μL of cold PBS containing 2% fetal calf serum. The first set of cells was stained with V450-conjugated anti-CD45 (BD Bioscience), BB515-conjugated anti-major histocompatibility complex II (MHCII) (BD Bioscience), phycoerythrin-conjugated anti-CD11c (BD Bioscience) for the detection of activated DCs, anti-CD11b+/F4/80+ for macrophages, and anti-CD11b+/Ly6G+ for neutrophils. The other set of cells was stained with peridinin chlorophyll protein complex-conjugated anti-CD3 (BD Bioscience), fluorescein isothiocyanate-conjugated anti-CD4 (eBiosience, San Diego, CA, USA), APC-conjugated anti-CD8, and phycoerythrin-conjugated anti-Foxp3 for the detection of T cells. The cells were analyzed by flow cytometry (LSR II; BD Bioscience). Flow cytometry was performed using an BD FACSCantoIIand analyzed by FlowJo software.
Bb515 conjugated anti major histocompatibility complex 2 mhcii
Manufactured by BD
BB515-conjugated anti-major histocompatibility complex II (MHCII) is a laboratory reagent that binds to the MHCII protein complex on the surface of cells. It is conjugated with the BB515 fluorescent dye, allowing for detection and analysis of MHCII-expressing cells using flow cytometry or other fluorescence-based techniques.
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2 protocols using bb515 conjugated anti major histocompatibility complex 2 mhcii
1
Multiparametric Analysis of Murine Splenocytes
2
Identification of Activated Dendritic Cells and Regulatory T Cells
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For the analysis, harvested cells from each mouse were distributed into 2 sets and were suspended in 50 μL of cold PBS containing 2% fetal calf serum. The first set of cells was stained with V450-conjugated anti-CD45 (BD Bioscience), BB515-conjugated anti-major histocompatibility complex II (MHCII) (BD Bioscience), phycoerythrin-conjugated anti-CD11c (BD Bioscience) and anaphase-promoting complex (APC)-conjugated anti-CD86 (BD Bioscience) for the detection of activated DCs. The other set of cells was stained with peridinin chlorophyll protein complex-conjugated anti-CD3 (BD Bioscience), fluorescein isothiocyanate-conjugated anti-CD4 (eBiosience, San Diego, CA, USA), APC-conjugated anti-CD25, and phycoerythrin-conjugated anti-Foxp3 for the detection of regulatory T cells. The cells were analyzed by flow cytometry (LSR II; BD Bioscience). The CD11c+MHCIIhigh cells were considered as migratory DCs (Supplementary Fig. S1 ).9 (link)
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