Nkp46 apc

Manufactured by BioLegend

NKp46-APC is a fluorochrome-conjugated monoclonal antibody that specifically binds to the NKp46 receptor, which is expressed on the surface of natural killer (NK) cells. NKp46 is a key activating receptor involved in the cytotoxic function of NK cells. The APC fluorochrome allows for the detection and analysis of NKp46-positive cells using flow cytometry.

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Lab products found in correlation

Tissue dissociation kit, by Miltenyi Biotec (1 mentions) Foxp3 pe, by Thermo Fisher Scientific (1 mentions) Anti cd3 fitc, by BioLegend (1 mentions) Cd4 bv510, by BD (1 mentions) Anti cd4 pe cy7, by Thermo Fisher Scientific (1 mentions) Anti cd8 percp cy5, by Thermo Fisher Scientific (1 mentions) Anti cd44 pe, by Thermo Fisher Scientific (1 mentions) Anti cd62l apc, by Thermo Fisher Scientific (1 mentions) Cd8 bv421, by BD (1 mentions) Cd3 percp cy5, by BD (1 mentions) Cd45 fitc, by BD (1 mentions) B220 pe cy7, by BD (1 mentions) Cd49b pe, by BioLegend (1 mentions) Cd45 bv510, by BioLegend (1 mentions) Symphony flow cytometer, by BD (1 mentions) Cd4 buv395, by BD (1 mentions) Tcrβ percp cy5, by BioLegend (1 mentions) Efluor 780, by Thermo Fisher Scientific (1 mentions) Software, by FlowJo (1 mentions) Attune nxt acoustic focusing cytometer, by Thermo Fisher Scientific (1 mentions)

3 protocols using nkp46 apc

Analyzing Tumor-Specific Immune Responses

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To systematically investigate the in vivo antitumour immune responses against mimic distant tumours, the tumours were harvested and treated with the tissue dissociation kit (Miltenyi Biotec, Germany) to produce a single-cell suspension according to the specified procedures. The harvested cells were further stained with several fluorochrome-conjugated antibodies: CD45-FITC (BD, Catalog: 561088), CD3-PerCP-Cy5.5 (BD, Catalog: 551163), CD4-BV510 (BD, Catalog: 563106), CD8-BV421 (BD, Catalog: 563898), NKp46-APC (Biolegend, Catalog: 137608), B220-PE-Cy7 (BD, Catalog: 552772) and Foxp3-PE (eBioscience, Catalog: 12–4771) and then analysed by FCM. For the analysis of the memory T cells, spleen cells of mice were harvested and stained with anti-CD3-FITC (Biolegend, Catalog: 100306), anti-CD4-PE-Cy7 (eBioscience, Catalog: 25–0041), anti-CD8-PerCP-Cy5.5 (eBioscience, Catalog: 45–0081), anti-CD44-PE (eBioscience, Catalog: 12–0441) and anti-CD62L-APC (eBioscience, Catalog: 17–0621) antibodies and then analysed by FCM. All antibodies were diluted ~100 times.

Yue W., Chen L., Yu L., Zhou B., Yin H., Ren W., Liu C., Guo L., Zhang Y., Sun L., Zhang K., Xu H, & Chen Y. (2019). Checkpoint blockade and nanosonosensitizer-augmented noninvasive sonodynamic therapy combination reduces tumour growth and metastases in mice. Nature Communications, 10, 2025.

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Multiparametric Flow Cytometry Protocol

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Single-cell suspensions were incubated in anti-CD16/CD32 Fc block antibody (5 μg/ml; clone 2.4G2; cat # 553141, BD Biosciences, San Jose, CA) and fixable viability dye eFluor 780 (Invitrogen, Carlsbad, CA) for 15 min at 4 °C in cold PBS. Cells were then washed once and stained with combinations of the following antibodies: CD45-BV510 (2 μg/ml; clone 30-F11; cat # 103138, BioLegend, San Diego, CA), TCRβ-PerCP-Cy5.5 (2 μg/ml; clone H57-597; cat # 109228, BioLegend, San Diego, CA), CD4-BUV395 (2 μg/ml; clone GK1.5; cat # 563790, BD Biosciences, San Jose, CA), CD8-af700 (5 μg/ml; clone KT15; cat # MCA609A700, Bio-Rad, Hercules, CA), NKp46-APC (1 μg/ml; clone 29A1.4; cat # 137607, BioLegend, San Diego, CA), CD49b-PE (2 μg/ml; clone DX5; cat # 108908, BioLegend, San Diego, CA), B220-BUV661 (2 μg/ml; clone RA3-6B2; cat # 612972, BD Biosciences, San Jose, CA), and Thy1.2-BUV805 (1 μg/ml; clone 53-2.1; cat # 741908, BD Biosciences, San Jose, CA). Data were acquired on a Symphony flow cytometer (BD Biosciences, San Jose, CA) and analyzed using FlowJo software (FlowJo LLC, Ashland, OR). Gating strategies for the identification of CD8⁺ T cells and NK cells are provided in Supplementary Fig. 13.

Taraborrelli L., Şenbabaoğlu Y., Wang L., Lim J., Blake K., Kljavin N., Gierke S., Scherl A., Ziai J., McNamara E., Owyong M., Rao S., Calviello A.K., Oreper D., Jhunjhunwala S., Argiles G., Bendell J., Kim T.W., Ciardiello F., Wongchenko M.J., de Sauvage F.J., de Sousa e Melo F., Yan Y., West N.R, & Murthy A. (2023). Tumor-intrinsic expression of the autophagy gene Atg16l1 suppresses anti-tumor immunity in colorectal cancer. Nature Communications, 14, 5945.

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Multiparameter Flow Cytometry Analysis

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Samples were run on an Attune NxT Acoustic Focusing Cytometer (Life Technologies). Analysis was performed in FlowJo. Gating was first done on forward scatter and side scatter to exclude debris. Doublets were excluded by gating on FSC area versus FSC height (Supplementary Fig. S8) Zombie Violet (BioLegend) was used to exclude dead cells from analyses. Antibodies: CD3-AF488 (BioLegend, clone HIT3a), Cd8a-AF488 (BioLegend, clone 53*6.7), CD45-PerCP/Cy5.5 (BioLegend, clone GK1.5), CD45-APC (BioLegend, clone 30-F11), NK1.1-APC (BioLegend, clone PK136), NKp46- APC (BioLegend, clone 29A1.4), CD11b- PE/Cy7 (BioLegend, clone M1/70), CD11c-AF488 (BioLegend, N418), IA/IE-AF700 (BioLegend M5/114.15.2), CD27 (BioLegend, clone LG.3A10), H-2- PE (BioLegend, clone M1/42), NKG2A-PE (Miltenyi Biotec, clone REA1161), H2-Dd-PE (BioLegend, clone 34-2-12), Qa-1b(b)-PE (BD Biosciences, clone 6A8.6F10.1A6), PD-L1–APC (BioLegend, clone 29E.2A3), CD49b-APC/Cy7 (BioLegend, clone DX5), and CD107b-FITC (BioLegend, clone 1D4B), IFNγ-PE (BioLegend, clone XMG1.2).

Taylor B.C., Sun X., Gonzalez-Ericsson P.I., Sanchez V., Sanders M.E., Wescott E.C., Opalenik S.R., Hanna A., Chou S.T., Van Kaer L., Gomez H., Isaacs C., Ballinger T.J., Santa-Maria C.A., Shah P.D., Dees E.C., Lehmann B.D., Abramson V.G., Pietenpol J.A, & Balko J.M. (2023). NKG2A Is a Therapeutic Vulnerability in Immunotherapy Resistant MHC-I Heterogeneous Triple-Negative Breast Cancer. Cancer Discovery, 14(2), 290-307.

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