Anti cd80 fitc

Manufactured by BD

Sourced in United States

Anti-CD80-FITC is a fluorescently labeled monoclonal antibody that binds to the CD80 (B7-1) antigen. CD80 is a cell surface protein expressed on the surface of antigen-presenting cells, including B cells, dendritic cells, and macrophages. The FITC (fluorescein isothiocyanate) label allows for the detection and analysis of CD80-positive cells using flow cytometry or other fluorescence-based techniques.

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Lab products found in correlation

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Close spelling variants of this product

CD80-FITC (43 citations) Anti-CD80 (43 citations) FITC-conjugated anti-CD80 (4 citations) FITC-labeled anti-CD80 (3 citations) FITC-conjugated anti-mouse CD80 (3 citations) Mouse anti-human CD80-FITC (2 citations) FITC-conjugated anti-human CD80 (2 citations)

29 protocols using anti cd80 fitc

Flow Cytometric Analysis of Human and Murine Dendritic Cells

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The following Abs were purchased from Becton Dickinson (San Diego, CA, USA; all diluted to 1:50): human phycoerythrin (PE) anti-CD11c, fluorescent isothiocyanate (FITC) anti-major histocompatibility complex (MHC) class II, FITC anti-CD80, FITC anti-CD86, PE anti-IL-3 receptor a chain (CD123), peridinin chlorophyll protein anti-human leukocyte antigen (HLA)-DR, FITC lineage cocktail 1 [lin1, contains monoclonal Abs CD3 (T cells), CD14 (monocytes/macrophages), CD16 (natural killer cells), CD19 (B cells), and CD56 (natural killer cells)], mouse FITC anti-MHC class II, FITC anti-CD80, FITC anti-CD86, and PE anti-CD11c.
Human peripheral blood cells were analyzed by three-color flow cytometry as reported previously^{24 (link)}. Human mDCs and pDCs were defined as Lin1⁻HLA-DR⁺/CD11c⁺ and Lin1⁻HLA-DR⁺/CD123⁺, respectively. Murine splenic DCs were identified as CD11c⁺ cells and the surface markers were incubated with anti-mouse MHC-II and CD86, respectively. Flow cytometry was performed on a FACS Caliber flow cytometer and analyzed with CellQuest Pro software (Becton Dickinson).

Ye Z., Zhong L., Zhu S., Wang Y., Zheng J., Wang S., Zhang J, & Huang R. (2019). The P-selectin and PSGL-1 axis accelerates atherosclerosis via activation of dendritic cells by the TLR4 signaling pathway. Cell Death & Disease, 10(7), 507.

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Modulating Cytokine Profiles in Dendritic Cells

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BMDCs (2×10⁶ cells) were transfected with IL-10 siRNA (100 nM, 300 nM, or 500 nM) in PINs for 24 hours at 37°C. Then, the siRNA-transfected DCs were stimulated with 5 µg/mL CpG ODNs in PCNs for 24 hours at 37°C. The expression of tumor necrosis factor (TNF)-α, IL-6, IL-12p70, and IL-10 in the DCs was analyzed using cytokine-specific enzyme-linked immunosorbent assays (ELISAs) (BD Biosciences, San Diego, CA, USA) according to the manufacturer’s instructions. Cytokine concentrations were quantified using a VICTOR3™ microplate reader (PerkinElmer, Inc., Waltham, MA, USA) by measuring the optical density (OD) at 450 nm and at the reference wavelength 570 nm. The expression of DC maturation markers was measured via flow cytometry after staining with FITC-anti-CD40, FITC-anti-CD80, and FITC-anti-CD86 antibodies (BD Biosciences).

Heo M.B., Kim S.Y., Yun W.S, & Lim Y.T. (2015). Sequential delivery of an anticancer drug and combined immunomodulatory nanoparticles for efficient chemoimmunotherapy. International Journal of Nanomedicine, 10, 5981-5993.

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Multicolor Flow Cytometry Immunophenotyping

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The analysis was performed by using a BD FACSCanto cytometer and BD FACSDiva software. APC anti-CD1a (# 559775), FITC anti-CD1a (#555806), BV421 anti-CD14 (#563743), FITC anti-CD14 (#555397), PE anti-CD14 (555398), PE anti-CD16 (#347617), FITC anti-CD64 (#555527), APC anti-CD209 (#551545), PE anti-CD1c (#564900), PE anti-CD86 (# 555658), FITC anti-CD80 (#560926), FITC anti-HLA-DR (#555811), APCCy7 anti-HLA-DR (#335796), APCCy7 anti-CD11b (#557754), APC anti-CD11c (#559877) were obtained from BD Biosciences. Sorting of CD1a + CD16 -cells was performed using a BD FACSCanto cytometer.

Erra Díaz F., Ochoa V., Merlotti A., Dantas E., Mazzitelli I., Gonzalez Polo V., Sabatté J., Amigorena S., Segura E, & Geffner J. (2020). Extracellular Acidosis and mTOR Inhibition Drive the Differentiation of Human Monocyte-Derived Dendritic Cells. Cell reports, 31(5).

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Eosinophil Surface Marker Analysis

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To confirm the presence of bona-fide bone marrow-derived eosinophils and their expression of various surface molecules upon T. canis stimulation, cutting-edge image flow cytometric analyses were performed for staining with the monoclonal antibodies: PE anti-Siglec-F⁺, FITC anti-MHC-II, FITC anti-CD69, APC anti-CD86, and FITC anti-CD80 (all purchased from BD Pharmingen). Samples were acquired using ImageStream Mark II and analyzed using Ideas software version 6.1 (both from Millipore-Sigma, Billerica, MA, USA).

Rodolpho J.M., Camillo L., Araújo M.S., Speziali E., Coelho-dos-Reis J.G., Correia R.D., Neris D.M., Martins-Filho O.A., Teixeira-Carvalho A, & Anibal F.D. (2018). Robust Phenotypic Activation of Eosinophils during Experimental Toxocara canis Infection. Frontiers in Immunology, 9, 64.

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Phenotypic Characterization of Murine Splenocytes

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Mice were immunized i.p. with 25 μg/animal of CpG ODN 1826 (InvivoGen) or with 5 μg/animal of Salmonella flagellin (FliC). 6 h after immunization, mice were euthanized and splenocytes were labeled. Fc receptors were blocked with Fc Block (BD Biosciences) and subsequently stained first with anti-CD19-Biotin (clone 1D3), anti-CD3-Biotin (clone 145.2C11), and anti-CD49b-Biotin (clone DX5) for 40 min on ice. After two washes with PBS-2% FBS, cells were then incubated anti-MHCII (I-A/I-E)-Alexa Fluor 700 (clone M5/114.15.2), anti-CD11c-BV421 (clone N418), anti-CD11b-PE.Cy7 (clone M1/70), anti-CD8α-BV786 (clone 52–67), anti-CD80-FITC (clone 16-10A1), anti-CD86-APC (clone GL1), anti-CD40-PE (clone 1C10), Streptavidin APC.Cy7 (all antibodies and the streptavidin were purchased from BD Biosciences) and Live and Dead Aqua (Life Technologies). Flow cytometry was performed using LSRFortessa (BD Biosciences) and results were analyzed in FlowJo software (version 9.3, Tree Star, San Carlo, CA, USA).

Antonialli R., Sulczewski F.B., Amorim K.N., Almeida B.D., Ferreira N.S., Yamamoto M.M., Soares I.S., Ferreira L.C., Rosa D.S, & Boscardin S.B. (2017). CpG Oligodeoxinucleotides and Flagellin Modulate the Immune Response to Antigens Targeted to CD8α+ and CD8α− Conventional Dendritic Cell Subsets. Frontiers in Immunology, 8, 1727.

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Analysis of NK Cell Surface Markers

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Cells were harvested after 24 h, 48 h, and 1 week. The expression of surface markers was analyzed by flow cytometry using anti-CD80-FITC, anti-CD70-PE, anti-CD40-PE and their corresponding isotype controls, and anti-CD56-FITC, anti-CD3-APC-Cy7 or anti-CD3-V500, anti-CD69-PE, anti-CD25-BV421 or anti-CD25-PE, and anti-CD54-APC or anti-CD54-PE (all from BD Biosciences) as recently described.^{33 (link)} Immunofluorescence was measured using a FACS Canto II (BD Biosciences), data were acquired with FACSDiva software (BD Biosciences) and evaluated with FCS Express software, version 5 (DeNovo Software). An average of approximately 6500 NK cells per measurement was acquired, with a minimum of 500 and a maximum of 23,000 cells.

Bosch N.C., Voll R.E., Voskens C.J., Gross S., Seliger B., Schuler G., Schaft N, & Dörrie J. (2019). NF-κB activation triggers NK-cell stimulation by monocyte-derived dendritic cells. Therapeutic Advances in Medical Oncology, 11, 1758835919891622.

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MHCII Cells Analysis in Stromal Visceral Fat

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The stromal visceral fat fraction (SVF) and spleen were used for the analysis of major histocompatibility complex class II (MHCII) cells. For this purpose, the obtained fraction was stained with anti-MHCII-PercpCy5.5 (562363 BD), anti-CD11C-PE (117308 BioLegend), anti-CD80-FITC (553768 BD), and anti-CD11b-APC (553312 BD); washed; and resuspended in PBS-5% FBS. The samples were analyzed using a FACSCanto II analyzer (BD Bioscience). Data were analyzed using FlowJo software for flow cytometry analysis.

Vecchiola A., Fuentes C.A., Solar I., Lagos C.F., Opazo M.C., Muñoz-Durango N., Riedel C.A., Owen G.I., Kalergis A.M, & Fardella C.E. (2020). Eplerenone Implantation Improved Adipose Dysfunction Averting RAAS Activation and Cell Division. Frontiers in Endocrinology, 11, 223.

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Dendritic Cell Functional Assay

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The BMDCs were obtained in the same way as in Section 2.5, and 1 × 10⁶ mL⁻¹ cells were cultured in 6-well plates (Thermo Fisher Scientific, Inc., Waltham, MA) at 37 °C in a humidified 5% CO₂ incubator. A blank control group and a LPS group as a positive control group (LPS group; the BMDCs were challenged using 5 µg/mL LPS) were designed for comparison. The DCs were cultured in an incubator under suitable growth conditions for 7 d and the medium was changed every 2 d. Then, the CYPPs were added to the DCs and incubated for 44 h. The DCs were collected and washed twice, then stained using anti-CD11c-APC, anti-MHC-II-PE-Cy5.5, anti-CD80-FITC, and anti-CD86-PE (BD Biosciences) in the dark at 4 °C for 30 min, which was in accordance with the protocol provided by the manufacturer. The cell pellets were dissociated by gentle pipetting for analysis using flow cytometry (BD FACSCalibur, Biosciences, Bedford, MA).

Luo L., Qin T., Huang Y., Zheng S., Bo R., Liu Z., Xing J., Hu Y., Liu J, & Wang D. (2017). Exploring the immunopotentiation of Chinese yam polysaccharide poly(lactic-co-glycolic acid) nanoparticles in an ovalbumin vaccine formulation in vivo. Drug Delivery, 24(1), 1099-1111.

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In Vivo Analysis of DC Maturation

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To examine DC maturation in vivo, TRAMP-C1 tumor-bearing C57BL/6 mice were treated with various strategy as described in in vivo therapy section. (n = 6) The inguinal lymph nodes were harvested 8 days post third treatment. The frequency of DC maturation in the LNs was then examined by flow cytometry after immunofluorescence staining with anti-CD11c-APC, anti-CD80-FITC, and anti-CD86-PE antibodies (BD Pharmingen™). To determine the intra-tumoral infiltration of CD8⁺(CD3⁺CD4⁻CD8⁺), lymphocytes were stained with anti-CD3-Brillant violet 510, anti-CD4-PE, anti-CD8-PE-cy7. All antibodies were diluted with the ratio of 1:100. And, signal color beads (OneComp eBeads™ Compensation Beads) and unstained bead were used for compensation. The frequency of CD8⁺(CD3⁺CD4⁻CD8⁺) lymphocytes in the LNs were also examined. Gating strategies used for cell sorting was shown in Supplementary Fig. 17.

Yu B., Choi B., Li W, & Kim D.H. (2020). Magnetic field boosted ferroptosis-like cell death and responsive MRI using hybrid vesicles for cancer immunotherapy. Nature Communications, 11, 3637.

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Characterization of DC Maturation Markers

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Mature DCs were electroporated without or with IKKβ-RNA as described above and harvested 24 h, 48 h, or 72 h after transfection. Cells were stained at 4°C in FACS buffer for 30 min with the following antibodies: anti-CD40-FITC (BD), anti-CD40-PE (BD), anti-CD25-FITC (Cymbus Technologies, Southampton, Hampshire, United Kingdom or BD), anti-CD25-PE (BD), anti-CD70-PE (BD), anti-OX40L-PE (BD), anti-CD80-FITC (BD), anti-CD83-PE (Miltenyi), anti-CCR7-FITC (R&D Systems), anti-CD86-FITC (Cymbus Technologies), anti-CD86-PE (Miltenyi, BD), and anti-PD-L1-PE (eBioscience) and with matched isotype controls: IgG1-FITC (BD, Miltenyi), IgG1-PE (Miltenyi), IgG2a-FITC (BD), IgG3-PE (eBioscience). The cells were then washed once with FACS buffer and were taken up in FACS buffer or a mixture of equal amounts of FACS-Fix (DPBS with 2% formaldehyde) and FACS buffer. Afterwards, the immunofluorescence was determined using a FACScan cytofluorometer equipped with Cell Quest software (BD). The DCs were discriminated based on their size and granularity by gating in the forward and side scatter channels. The specific mean fluorescence intensities (specific MFI) were calculated by subtraction of the background mean fluorescence intensity obtained with the isotype control antibodies. All values were set in relation to the 24 h control condition to calculate the fold induction.

Gerer K.F., Erdmann M., Hadrup S.R., Lyngaa R., Martin L.M., Voll R.E., Schuler-Thurner B., Schuler G., Schaft N., Hoyer S, & Dörrie J. (2017). Preclinical evaluation of NF-κB-triggered dendritic cells expressing the viral oncogenic driver of Merkel cell carcinoma for therapeutic vaccination. Therapeutic Advances in Medical Oncology, 9(7), 451-464.

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