Anti mouse nk1.1 apc

Manufactured by BioLegend

Anti-mouse NK1.1-APC is a fluorochrome-conjugated monoclonal antibody that binds to the NK1.1 antigen expressed on natural killer cells and a subset of T cells in mice.

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3 protocols using anti mouse nk1.1 apc

Multiparametric Flow Cytometry Analysis

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For surface staining, immune cells in skin tissues were first blocked with anti-mouse CD16/32 (101302, Biolegend) for 30 min at 4°C in PBS containing 1% FBS. After washing, cells were stained with following antibodies: anti-mouse TCRβ-AF700(109224, Biolegend), anti-mouse TCR γ/δ-FITC antibody (118106, Biolegend), anti-mouse CD8α-PE (100708, Biolegend), anti-mouse CD11c-PE (117308, Biolegend), anti-mouse F4/80-AF488 (123120, Biolegend), anti-mouse NK1.1-APC(108710, Biolegend), anti-mouse B220-Alexa Fluor 700 (103232, Biolegend), anit-mouse Ly6C (128026, Biolegend), anti-mouse I-A/I-E-BV605 (107639, Biolegend), anti-mouse CD3ε-BUV737 (612771, BD Bioscience), anti-mouse CD11b-BUV737 (612800, BD Bioscience; For intracellular staining, cells were washed with PBS, fixed and permeabilized with Intracellular Staining Buffer Set (00-5523-00, Invitrogen) according to the manufacturer’s protocol. Cells were stained for 1 h at 4°C with different antibodies: anti-mouse TNFα-BV605 (506329, Biolegend), anti-mouse IL-17A-Pacific blue (506918, Biolegend), anti-mouse IL-36α-APC (32103-05161, Assaypro), anti-mouse IL-36γ (PAL621Mu01, Cloud-clone Corp), or control antibody (goat anti-rabbit IgG, 111-136-144, Jackson ImmunoResearch). Cells were acquired either by BD FACS Fortessa or BD FACS Aria II Cell Sorter. All the data were analyzed using FlowJo 10.1.

Tamaru Y, & Doi R.H. (1999). Three Surface Layer Homology Domains at the N Terminus of the Clostridium cellulovorans Major Cellulosomal Subunit EngE. Journal of Bacteriology, 181(10), 3270-3276.

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Multiparametric Flow Cytometry Analysis

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Hao J., Jin R., Zeng J., Hua Y., Yorek M.S., Liu L., Mandal A., Li J., Zheng H., Sun Y., Yi Y., Yin D., Zheng Q., Li X., Ng C.K., Rouchka E.C., Egilmez N.K., Jabbari A, & Li B. (2022). Consumption of fish oil high-fat diet induces murine hair loss via epidermal fatty acid binding protein in skin macrophages. Cell reports, 41(11), 111804.

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Phenotypic Characterization of Immune Cells

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After incubation with CPP-conjugated molecules, the cells were resuspended in 100 μl antibody mixture diluted in PBS and incubated at 4°C for 15 min. To discriminate each cell subset from the mixed cell population, different antibody combinations were used. For cell type analysis, anti-mouse CD4-PerCP-Cy5.5 (BioLegend, Clone:RM4-5), anti-mouse CD19-PE-Cy7 (BioLegend, Clone:6D5), anti-mouse NK1.1-APC (BioLegend, Clone:PK136), anti-mouse CD11b-PE-Cy7 (eBioscience, Clone:H57-597), anti-mouse CD11c-APC (eBioscience, Clone:N418), anti-mouse I-A/I-E-APC-Cy7 (BioLegend, Clone:M5/114.15.2), and anti-mouse F4/80-PerCP-Cy5.5 (BioLegend, Clone:BM8) were used. Also, anti-mouse CD16/32 (BioLegend, Clone:93) was used for blocking the Fc receptor. For activated T cell analysis, anti-mouse CD69-FITC (BioLegend, Clone:H1.2F3), anti-mouse CD4-APC (eBioscience, Clone:GK1.5), and anti-mouse CD8-PerCP-Cy5.5 (eBioscience, Clone:53–6.7) were used. For activated B cell analysis, anti-mouse CD86-PE-Cy5 (BioLegend, Clone:GL-1), anti-mouse IgG2a, kappa-isotype (BioLegend, Clone:RTK2758), and anti-mouse CD19-PE (BioLegend, Clone:6D5) were used.

Lim S., Lee J.A., Koo J.H., Kang T.G., Ha S.J, & Choi J.M. (2016). Cell Type Preference of a Novel Human Derived Cell-Permeable Peptide dNP2 and TAT in Murine Splenic Immune Cells. PLoS ONE, 11(5), e0155689.

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