Nk1.1 bv421

Manufactured by BioLegend

Sourced in United States

NK1.1-BV421 is a monoclonal antibody that binds to the NK1.1 antigen, which is expressed on natural killer (NK) cells and a subset of T cells in mice. The antibody is conjugated to the Brilliant Violet 421 fluorescent dye, which can be used for flow cytometric analysis of NK1.1-positive cell populations.

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2 protocols using nk1.1 bv421

Comprehensive Immune Cell Profiling

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The adipose SVCs and splenocytes were incubated for 10 minutes with CD16/32 antibody (FC block) (eBioscience) and then stained with other fluorochrome-conjugated antibodies for 20 minutes at 4℃. All cells were stained with antibodies against CD45-PE/Cy7 (eBioscience), CD4-BV500 (BD Biosciences), CD8-APC/Cy7 (Biolegend), CD19-PE (Biolegend), NK1.1-BV421 (Biolegend), NKp46-PE/Dazzle594 (Biolegend), CD11b-BV786 (Biolegend), CD11c-APC (Biolegend), CD3-BV711 (Biolegend), and F4/80-BV650 (Biolegend). 7AAD (Invitrogen) was used for live/dead staining. The stained cells were washed and analyzed with an LSRFortessa flow cytometer (BD Biosciences). NK cells were defined as CD45⁺ F4/80⁻ CD19⁻ CD3⁻ NK1.1⁺ in lymphocyte gating. The NKp46⁺ and GFP⁺ cells were then gated. ATMs were defined as CD45⁺ CD19⁻ CD3⁻ NK1.1⁻ F4/80⁺ CD11b⁺. Total cell numbers were counted with a hemocytometer and normalized according to the tissue weight (cells/g). Flow cytometric data were analyzed by using the FlowJo software (Flow Jo).

Nathalie G., Bonamichi B.D., Kim J., Jeong J., Kang H., Hartland E.R., Eveline E, & Lee J. (2023). NK cell-activating receptor NKp46 does not participate in the development of obesity-induced inflammation and insulin resistance. Molecules and Cells, 47(3), 100007.

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Liver Cell Immunophenotyping by Flow Cytometry

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After obtain single cells of perfused liver, the cells were incubated with Fixable Viability Dye(BioLegend, San Diego, CA, USA) to distinguish between dead and living cells. After application of the FcR Blocking Reagent (Miltenyi Biotec, Bergisch Gladbach, Germany), single-cell suspensions were stained with the following fluorochrome-conjugated antibodies including CD45-Super bright 600 (eBioscience, San Diego, CA, USA), CD3-FITC, CD4-PerCP-Cy5.5, CD8-PE-Cy7, NK1.1-BV421, CD19-APC-Cy7 and FasL-APC (the rest of antibodies were BioLegend). Flow cytometry was performed using a BD FACS Canto II, and the results were analyzed using FlowJo 10.4 software (Tree Star, Ashland, OR, USA).

Zhang Y., Li J., Li H., Jiang J., Guo C., Zhou C., Zhou Z, & Ming Y. (2022). Single-cell RNA sequencing to dissect the immunological network of liver fibrosis in Schistosoma japonicum-infected mice. Frontiers in Immunology, 13, 980872.

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