Cd11b brilliantviolet bv 421

Manufactured by BioLegend

CD11b-BrilliantViolet (BV) 421 is a fluorochrome-conjugated antibody that binds to the CD11b cell surface antigen. CD11b is a subunit of the integrin Mac-1 (also known as complement receptor 3 or CR3) and is expressed on the surface of various immune cells, including monocytes, macrophages, and neutrophils. This product can be used for the detection and analysis of CD11b-expressing cells in flow cytometry applications.

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CD11b (457 citations)

2 protocols using cd11b brilliantviolet bv 421

Multiparameter Flow Cytometric Analysis of Bone Marrow

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BM cells from femur were flushed out and erythrocytes were lysed. The remaining cells were stained with eBioscience Fixable Viability Dye eFluor 780 (Thermo Fisher Scientific), followed by incubation with Fc-gamma receptor block (Becton Dickinson (BD)). Afterwards, the cells were stained with the following fluorescent-labeled antibodies (Biolegend): CD11b-BrilliantViolet (BV) 421, CD19-BV421, CD8-BV510, CD11c-FITC, RANK- Phycoerythrin (PE), RANKL-PE, F4/80-PE Cyanine7 (Cy7), CD3-PECy7, Gr-1-Peridinin-Chlorophyll-Protein (PerCP), MCSF-R-Allophycocyanin (APC), CD4-APC. Fluorescence minus one (FMO) stained samples were used as controls. The cells were either immediately acquired or fixed in 4% paraformaldehyde, before acquisition using FACSVerse (BD). The data were analyzed with the FlowJo software Version 10 (FlowJo).

Scheffler J.M., Grahnemo L., Engdahl C., Drevinge C., Gustafsson K.L., Corciulo C., Lawenius L., Iwakura Y., Sjögren K., Lagerquist M.K., Carlsten H., Ohlsson C, & Islander U. (2020). Interleukin 17A: a Janus-faced regulator of osteoporosis. Scientific Reports, 10, 5692.

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Multiparameter Flow Cytometry Identification

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Cells were stained in a staining buffer containing 2% heat-inactivated rat sera, 1 μg anti-mouse CD16/32 (clone: 93, BioLegend), and fluorochrome-conjugated antibodies in the dark for 20 minutes at 4°C. Eosinophils were identified using CD45 Alexa Fluor^® 700 (clone: 30-F11, BioLegend), CD11b Brilliant violet (BV) 421™ (clone: M1/70, BioLegend), Ly6C FITC (clone: HK1.4, BioLegend), Ly6G APC Cy7 (clone: 1A8, BioLegend), and Siglec-F PE (clone: S17007L, BioLegend). ILC2s were characterised using CD45 Alexa Fluor^® 700 (clone: 30-F11, BioLegend), lineage cocktail PE (CD3ϵ clone: 145-2C11, Ly-6G/Ly-6C clone: RB6-8C5, CD11b clone: M1/70, CD45R/B220 clone: RA3-6B2, TER-119 clone: Ter-119, BioLegend), IL-7Rα (CD127) PE Cy7 (clone: A7R34, BioLegend), ICOS APC (clone: C398.4A, BioLegend), ST2 (IL-33Rα) BV421™ (clone: DIH9, BioLegend), and KLRG1 BV510™ (clone: 2F1/KLRG1, BioLegend). 7-aminoactinomycin D (7-AAD) staining was used to identify non-necrotic (‘Live’) cells. Cells were acquired on an LSRFortessa (BD Biosciences) and FACS Aria I (BD Biosciences) for cell sorting. Single-stained and unstained controls were used to compensate for spectral overlap. Data were analyzed using FlowJo^© V10 (Treestar, Ashland, OR).

Chetty A., Darby M.G., Pillaye J., Taliep A., Cunningham A.F., O’Shea M.K., Katawa G., Layland L.E., Ritter M, & Horsnell W.G. (2023). Induction of Siglec-FhiCD101hi eosinophils in the lungs following murine hookworm Nippostrongylus brasiliensis infection. Frontiers in Immunology, 14, 1170807.

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