Cd80 clone 16 10a1

Manufactured by BioLegend

Sourced in United States

CD80 (clone 16-10A1) is a lab equipment product offered by BioLegend. It is a monoclonal antibody that binds to the CD80 protein, which is a costimulatory molecule expressed on antigen-presenting cells. The core function of this product is to facilitate the detection and analysis of CD80-expressing cells.

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Lab products found in correlation

6 protocols using cd80 clone 16 10a1

Detailed Immune Cell Profiling by Flow Cytometry

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Binding of the fusion protein was detected using PE-anti-human IgG Fc (clone HP6017,1 μg/ml). PD-L1 (clone 10 F.9G2, 1 μg/ml), IFNAR1 (clone MAR1-5A3, 1 μg/ml), CD45 (clone 30-F11, 0.5 μg/ml), CD80 (clone 16-10A1, 0.5 μg/ml), CD86 (clone GL1, 0.5 μg/ml) were from BioLegend or eBioscience. Cells suspended in FACS buffer (1% bovine serum albumin and 0.05% NaN3) were blocked with anti-CD16/32 (anti-FcγIII/ II receptor, clone 2.4G2) for 30 min and then stained with specific antibodies for 30 min on ice. Samples were analyzed on a FACS Calibur or Fortessa flow cytometer (BD Biosciences). Data were analyzed using FlowJo software (TreeStar).

Liang Y., Tang H., Guo J., Qiu X., Yang Z., Ren Z., Sun Z., Bian Y., Xu L., Xu H., Shen J., Han Y., Dong H., Peng H, & Fu Y.X. (2018). Targeting IFNα to tumor by anti-PD-L1 creates feedforward antitumor responses to overcome checkpoint blockade resistance. Nature Communications, 9, 4586.

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Multicolor Flow Cytometry Analysis

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Single-cell suspensions from spleens, dLNs, non-dLNs, or tumor tissues were prepared as previously described (42 (link)). Combinations of the following fluorochrome-conjugated antibody were used for cell surface or intracellular staining to define populations of NK, CD8, and subsets of CD4 T cells: CD3e (clone 145-2c11), CD8a (clone 53-6.7), CD4 (clone GK1.5), NK1.1 (clone PK136), NKG2D (clone CX5), CD44 (clone eBio4B10), CD11c (clone N418), MHCII (clone M5/114.15.2), CD80 (clone 16-10A1), CD86 (clone PO3), CD40 (clone 1C10), and CD3ζ (clone 6B10.2, BioLegend). For ex vivo restimulation, freshly isolated single-cell suspension was cultured in complete RPMI 1640 medium containing PMA (50 ng/ml) and ionomycin (500 ng/ml) for 6 hours before it was analyzed for IFN-γ production by intracellular staining with an antibody specific to IFN-γ (XMG1.2). Multicolored flow cytometry analyses were performed on LSR II (BD). Data were analyzed with FlowJo software (Tree Star).

Zhang J., Liu D., Li G., Staveley-O’Carroll K.F., Graff J.N., Li Z, & Wu J.D. (2017). Antibody-mediated neutralization of soluble MIC significantly enhances CTLA4 blockade therapy. Science Advances, 3(5), e1602133.

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Multiparameter Flow Cytometry Analysis

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MOC cells were harvested and non-specific binding was blocked with anti-CD16/32 antibodies (Biolegend) prior to staining. Tissues were prepared into single-cell suspensions as previously described (14 (link)), followed by anti-CD16/32 antibody staining. Cell surface staining was performed using flourophore conjugated anti-mouse CD45.2 clone 104, CD3 clone 145-2C11, CD4 clone GK1.5, CD8 clone 53-6.7, NK1.1 clone PK136, CD31 clone 390, PD-L1 clone 10F.9G2, H2-K^b clone AF6-88.5, CD107a clone 1D4B, CD69 clone H1.2F3, PD-1 clone RMP1-30, CD11b clone M1/70, CD11c clone N418, CD80 clone 16-10A1, and CD86 clone GL-1 antibodies from Biolegend, 41BB clone 17B5 and OX40 clone OX-86 from eBioscience, and CD8 clone KT15 from MBL (Woburn, MA). H2Kb:KSPWFTTL (p15E_604–611) tetramer was purchased from MBL. Dead cells were excluded via 7AAD negativity. Isotype control antibodies and a “fluorescence minus one” method of antibody combination were used for specific staining validation. Data was acquired on a FACSCanto using FACSDiva software (BD Biosciences) and analyzed on FlowJo software vX10.0.7r2.

Moore E., Clavijo P.E., Davis R., Cash H., Van Waes C., Kim Y, & Allen C. (2016). Established T-cell inflamed tumors rejected after adaptive resistance was reversed by combination STING activation and PD-1–pathway blockade. Cancer immunology research, 4(12), 1061-1071.

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Evaluating DC Modulation by BG Ionic Dissolution

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To determine the effect of the BG ionic dissolution products on DC modulation, the above mentioned DCs co-incubated with varying amounts of CCM containing IDPs from B3, B3-Cu, B3-Zn, and B3-Cu-Zn BGs were analyzed for the expression of DC specific surface markers using flow cytometry. The mature surface DC phenotype was analyzed using antibodies specific for CD11c (clone N418, BioLegend), MHCII (clone M5/114.15.2, BioLegend), CD86 (clone GL-1, BD Biosciences), CD80 (clone 16-10A1, BioLegend) and CD83 (clone Michel-19, BioLegend). Samples were stained using 7-AAD (7-aminoactinomycin D) to determine the toxicity of the substances and to perform the analysis of living cells (ThermoFisher Scientific). Samples were measured using the BD FACS Canto II cytometer and were analyzed using FCS Express 5 Flow Cytometry Software (DeNovo software).

Schuhladen K ., Stich L ., Schmidt J ., Steinkasserer A ., Boccaccini AR , & Zinser E . (2020). Cu, Zn doped borate bioactive glasses: antibacterial efficacy and dose-dependent in vitro modulation of murine dendritic cells. Biomaterials science, 8(8).

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Phenotypic Analysis of Co-stimulatory Molecules

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To detect the expression levels of co-stimulatory molecules on the cell surface of RAW-267.4 cells and BMDCs, fluorescein isothiocyanate (FITC)-conjugated rat anti-mouse CD40 (clone 3/23, Biolegend, San Diego, CA, USA), CD80 (clone 16-10A1, Biolegend, San Diego, CA, USA), CD86 (clone GL-1, Biolegend, San Diego, CA, USA), or MHC class II (I-A/I-E) mAbs (clone M5/114.15.2, Biolegend, San Diego, CA, USA) were used. In all stainings, IgG receptors on the cell surface were blocked using either monoclonal antibody (mAb) against CD16/CD32 (clone 2.4G2, Thermo Fisher Scientific, Waltham, MA, USA), or normal murine IgG antibodies (Santa Cruz Biotechnology, Dallas, TX, USA). BMDCs were additionally stained with allophycocyanin (APC)-conjugated anti-mouse CD11c mAb (clone N418, Biolegend, San Diego, CA, USA) to gate the DC population. Fluorescence intensity of stained cells (10,000 cell detection) was measured with flow cytometry using a FACSLyric (BD Bioscience, Franklin Lakes, NJ, USA). Data were analyzed using FlowJo V. 7 (BD Bioscience, Franklin Lakes, NJ, USA).

Cheng T.Y., Lin Y.J., Saburi W., Vieths S., Scheurer S., Schülke S, & Toda M. (2021). β-(1→4)-Mannobiose Acts as an Immunostimulatory Molecule in Murine Dendritic Cells by Binding the TLR4/MD-2 Complex. Cells, 10(7), 1774.

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Characterizing Monocyte Activation by CAR T Cells

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CD14⁺ monocytes were isolated from PBMCs by positive selection. Isolated monocytes (10,000 cells) were plated in a 96-wells flat-bottom plate. 24 hours later, 10,000 CAR T cells and 10,000 tumor cells (MM1.S) were added in each well and were incubated for 24 hours. After incubation, the wells were washed with phosphate-buffered saline twice and detached from the wells with Trypsin/EDTA solution (Lonza) for 20 minutes. Monocytes were stained with CD3 (clone SK7, BD Biosciences), CD14 (clone HCD14, BioLegend), HLA-DR (clone G46-6, BD Biosciences), CD86 (clone HA5.2B7, Beckman Coulter), and CD80 (clone 16-10A1, BioLegend) (staining with 1 μg per mL of sample, incubation for 30 minutes at 4°C in PBS with 1% BSA, 10% FBS and 0.1% NaN3 sodium azide). Flow cytometry was performed on BD LSR Fortessa.

Katsarou A., Sjöstrand M., Naik J., Mansilla-Soto J., Kefala D., Kladis G., Nianias A., Ruiter R., Poels R., Sarkar I., Patankar Y.R., Merino E., Reijmers R.M., Frerichs K.A., Yuan H., de Bruijn J., Stroopinsky D., Avigan D., van de Donk N.W., Zweegman S., Mutis T., Sadelain M., Groen R.W, & Themeli M. (2021). Combining a CAR and a chimeric costimulatory receptor enhances T cell sensitivity to low antigen density and promotes persistence. Science translational medicine, 13(623), eabh1962.

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