Bv421 anti cd161

Manufactured by BD

Sourced in United States

BV421-anti-CD161 is a fluorochrome-conjugated antibody that binds to the CD161 antigen. CD161 is a type II transmembrane receptor expressed on a subset of T cells and natural killer cells. This antibody can be used for the identification and analysis of these cell populations by flow cytometry.

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5 protocols using bv421 anti cd161

Multiparameter Immunophenotyping of Immune Cells

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PerCp-Cy5.5 anti-CD161, PE anti-RORγt, PE anti–γδ TCR, biotinylated anti–γδ TCR, PerCp-Cy5.5 anti-CD45RA, and APC anti–αβ TCR antibodies were from eBioscience. A647 anti-PLZF, APC-Cy7 anti-CD4, PE-Cy7 anti-CD8, BV421 anti-CD3, BV421 anti-CD161, PE anti-STAT4 (pY693), PE anti–IL-6R, PE anti–IL-12Rβ1, PE anti–IL-21R, PE mouse IgG1 isotype control, PerCP-Cy5.5 anti-STAT3 (pY705), and FITC anti–Vδ2 TCR antibodies were from BD. FITC anti–Vα24 TCR and PE and biotinylated anti-Vβ11 antibodies were from Beckman Coulter. FITC anti–Vδ1 TCR was from Thermo Fisher Scientific. APC or APC-Cy7 anti-Vα7.2 and PE-Cy7 anti–αβ TCR antibodies were from BioLegend. FITC anti-CCR7 antibody was from R&D Systems. MR1–5-OP-RU tetramers have been described previously (Reantragoon et al., 2013 (link); Corbett et al., 2014 (link)).

Wilson R.P., Ives M.L., Rao G., Lau A., Payne K., Kobayashi M., Arkwright P.D., Peake J., Wong M., Adelstein S., Smart J.M., French M.A., Fulcher D.A., Picard C., Bustamante J., Boisson-Dupuis S., Gray P., Stepensky P., Warnatz K., Freeman A.F., Rossjohn J., McCluskey J., Holland S.M., Casanova J.L., Uzel G., Ma C.S., Tangye S.G, & Deenick E.K. (2015). STAT3 is a critical cell-intrinsic regulator of human unconventional T cell numbers and function. The Journal of Experimental Medicine, 212(6), 855-864.

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Multiparametric Flow Cytometry Analysis

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Isolated PBMCs were stained in duplicate with FITC-anti-CD3 (BD Biosciences, San Diego, CA, USA), PerCP-Cy5.5-anti-CD19 (BD Biosciences), BV421-anti-CD161 (BD Biosciences), APC-anti-TCRVα7.2 (BioLegend, San Diego, CA, USA), PE-CF594-anti-TCRγδ (BD Biosciences), PE-Cy7-anti-CD8α (BD Biosciences), and PE-anti-CD8β (BD Biosciences) antibodies at 4 °C for 30 min in the dark. Isotype-matched control antibodies were used as negative controls. The frequencies of various T-cell subsets were determined by flow cytometry analysis using the FACSAria II (BD Biosciences) and FlowJo software (v7.6.2; TreeStar, San Carlos, CA, USA).

Zhao D., Zhong W., Han D., Li Y., Jiang Y, & Gu G. (2019). Elevated frequencies of total and MAIT cell subsets in patients with knee osteoarthritis. PeerJ, 7, e7443.

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Comprehensive Phenotyping of T Cell Subsets

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PBMC or cultured T cells were stained for 30 min at 4°C with following fluorochrome-conjugated Abs: APC-Cy7-anti-CD3, v500-anti-CD4, PE-Cy5-anti-CD25, APC-anti-CD25, PE-Cy7-anti-CD45RA, BV421-anti-CD161, (six from BD Bioscience), FITC-anti-GARP (Enzo Life Science, Farmingdale, NY), PE-Cy7-anti-GITR, APC-Cy7-anti-CD73, BV421-anti-CD39, PerCP-Cy5.5-anti-CTLA-4, PE-Cy7-anti-CXCR3 (five from BioLegend, San Diego, CA), PE-anti-IL-1RI, APC-anti-IL-1RII, and FITC-anti-IL-1RII (three from R and D systems). For intracellular cytokine staining (ICS), cultured T cells were re-stimulated for 6 hr with PMA (50 ng/ml) and ionomycin (1 μg/ml) in the presence of BFA for last 4 hr, followed by staining with PE-anti-IL-1RI and FITC-anti-IL-1RII Abs. The stained cells were fixed and permeabilized using Fix/Perm buffer (BioLegend), followed by staining with anti-IL-17A, anti-IFN-γ (both from eBioscience), and anti-Foxp3 (Biolegend) Abs. The cells were acquired using a BD LSRFortessa (BD Bioscience) and analyzed using FlowJo software (Tree Star, Ashland, OR).

Kim D.H., Kim H.Y., Cho S., Yoo S.J., Kim W.J., Yeon H.R., Choi K., Choi J.M., Kang S.W, & Lee W.W. (2021). Induction of the IL-1RII decoy receptor by NFAT/FOXP3 blocks IL-1β-dependent response of Th17 cells. eLife, 10, e61841.

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Identifying and Characterizing MAIT Cells and Monocytes in Liver Disease

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Mononuclear cells were stained with ethidium monoazide (EMA, Sigma-Aldrich), anti-CD161-BV421, anti-CD16-V500, anti-CD8-BV605, anti-CD19-PECy5, anti-CD107a-PE (BD Biosciences), anti-TCRVα7.2-FITC, anti-CD3-AlexaFluor700 and anti-CD14-PE-Cy7 (Biolegend) on the day they were isolated from blood samples and liver biopsies. Anti-CD69-APC-Cy7 (Biolegend), anti-HLA-DR-BV711 and anti-CD107a-PE (BD Biosciences) were included to assess activation (CD69, HLA-DR) and degranulation (CD107a).
MAIT cells were identified as CD3⁺CD161⁺TCRVα7.2⁺ cells and studied in 35 paired blood samples and 32 paired liver biopsies prior to and at week 4 of antiviral therapy. Monocytes were identified as SSC^highHLA-DR⁺CD14⁺ cells and studied in 34 paired blood samples and 29 paired liver biopsies prior to and at week 4 of therapy. Liver biopsies with less than 41 events in the MAIT cell gate were excluded from analysis for activation and degranulation markers. CD14⁺⁺CD16⁻, CD14⁺⁺CD16⁺ and CD14⁺CD16⁺⁺ monocyte subsets were identified as described ^{31 (link)} and analyzed for activation marker expression if their event counts exceeded the 25^th percentile of the respective subset in all liver biopsies (n=33, 33, 23 and 23 events, respectively).

Bolte F.J., O’Keefe A.C., Webb L.M., Serti E., Rivera E., Liang T.J., Ghany M, & Rehermann B. (2017). Intra-hepatic Depletion of Mucosal Associated Invariant T cells in Hepatitis C Virus-induced Liver Inflammation. Gastroenterology, 153(5), 1392-1403.e2.

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Multifaceted Immune Cell Analysis

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The isolated cells were stained with the fluorescent-labeled antibodies, including anti-CD3-PE-CF594 (UCHT1), anti-CD8-V500 (RPA-T8, BD Horizon), anti-CD161-BV421 (DX12), anti-CD4-PerCP-Cy5.5 (RPA-T4), anti-TCRγδ-PE (B1), anti-CD45RO-APC-H7 (UCHL1, BD PharMingen), anti-IL-18Rα-FITC (H44), anti-TCR Vα7.2-APC (3C10, Biolegend) and anti-CCR6-PE (R6H1, eBioscience). The stained cells were characterized on the FACS Aria II flow cytometers (BD Biosciences) and analyzed using the FlowJo software (TreeStar).

Ling L., Lin Y., Zheng W., Hong S., Tang X., Zhao P., Li M., Ni J., Li C., Wang L, & Jiang Y. (2016). Circulating and tumor-infiltrating mucosal associated invariant T (MAIT) cells in colorectal cancer patients. Scientific Reports, 6, 20358.

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