Anti cd86 fitc

Manufactured by Thermo Fisher Scientific

Sourced in United States

Anti-CD86-FITC is a fluorochrome-conjugated antibody that binds to the CD86 cell surface marker. CD86 is a costimulatory molecule expressed on antigen-presenting cells. This product can be used for the identification and enumeration of CD86-positive cells in flow cytometry applications.

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Close spelling variants of this product

Anti-CD86 (32 citations) CD86-FITC (24 citations) FITC-conjugated anti-CD86 (5 citations) Anti-mouse CD86-FITC (5 citations) FITC-conjugated anti-mouse CD86 (4 citations) FITC‐conjugated anti‐mouse CD86 antibody (3 citations) FITC-labeled anti-mouse CD86 (3 citations) FITC-anti-CD86 clone GL1 (2 citations) FITC-conjugated anti-mouse CD80/CD86/CD4 (2 citations) Anti-CD86-FITC antibodies (2 citations)

15 protocols using anti cd86 fitc

Immunomodulatory effects of ALX in EAE

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The harvested BMDC (2 × 10⁶ /well) were stimulated in triplicate with LPS (1 μg/mL), alone or together with ALX at 2 or 10 μM in 6-well plates for 2 days. The cells were stained with PE-anti-CD11c, Percp-cy5.5-anti-MHC-II, FITC-anti-CD80 or FITC-anti-CD86, or isotype controls (eBioscience), washed, and analyzed by flow cytometry.
On days 24–26 post-immunization, six mice from each group were sacrificed, and their splenic mononuclear cells were isolated. The cells were stimulated with Cell Stimulation Cocktail (PMA + Ionomycin, eBioscience) for 5 h. The cells were stained with FITC-anti-CD4 (eBioscience), fixed, and permeabilized. After being washed, the cells were stained intracellularly with PE-anti-IFN-γ or PE-anti-IL-17 (eBioscience). The frequency of Th1 (CD4⁺IFN-γ⁺) and Th17 (CD4⁺Th17⁺) cells in total CD4⁺ T cells was determined by flow cytometry.
On days 15 post-immunization (the early stage of EAE), splenic mononuclear cells were isolated from the vehicle or ALX-treated EAE mice. The cells were stimulated with 20 μg/ml of MOG35-55 for 48 h and stained with PE-anti-CD11c, Percp-cy5.5-anti-MHC-II, FITC-anti-CD80 or FITC-anti-CD86, or isotype controls (eBioscience), and their expression levels were characterized by flow cytometry.

Quan M.Y., Song X.J., Liu H.J., Deng X.H., Hou H.Q., Chen L.P., Ma T.Z., Han X., He X.X., Jia Z, & Guo L. (2019). Amlexanox attenuates experimental autoimmune encephalomyelitis by inhibiting dendritic cell maturation and reprogramming effector and regulatory T cell responses. Journal of Neuroinflammation, 16, 52.

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Characterization of Mast Cell Phenotype

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Following the blocking of Fcγ receptors with 2.4G2 (BD Pharmingen, Franklin Lakes, NJ, USA), cells were stained with PE-anti FcεRI (eBioscience), APC-anti FcεRI (eBioscience), FITC-anti KIT (BD Pharmingen), PE-anti KIT (BD Pharmingen), FITC-anti MHC clssII (I-Ad) (eBioscience), FITC-anti-CD40 (eBioscience), FITC-anti- CD80 (eBioscience), FITC-anti- CD86 (eBioscience), FITC-anti-PD-L1(eBioscience), APC-anti-CD11c (eBioscience) antibody to examine the expression of FcεRI, KIT, MHC class II, CD40, CD80, CD86, and PDL-1. To measure cytoplasmic protein, the cells were fixed and treated with BD Cytofix/Cytoperm™ Plus Fixation/Permeabilization Kit with BD GolgiPlug (BD Bioscience) according to the manufacturer's protocol. The cells were then incubated for 1h at 4°C. After washing with PBS, cells stained with Ab were analyzed using a FACS Calibur flow cytometer (BD Biosciences) and associated CellQuest software.

Ito T., Egusa C., Maeda T., Numata T., Nakano N., Nishiyama C, & Tsuboi R. (2015). IL-33 promotes MHC class II expression in murine mast cells. Immunity, Inflammation and Disease, 3(3), 196-208.

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Phenotypic Modulation of Dendritic Cells by Botanical Compound

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To assess the effects of BC on immune cells, D1DCs (a murine dendritic cell line) [22 (link)] were cultured with increasing amounts of BC extracts (up to 20 mg/mL). After 48 h, the D1DCs were detached using PBS/EDTA, washed with FACS buffer (PBS with 0.5% BSA and 0.02% sodium azide) and stained at 4 °C for 30 min with the following antibodies: anti-CD40-APC (Biolegend, San Diego, CA, USA, clone 3/23, #12-4612), anti-CD86-FITC (eBioscience, San Diego, CA, USA) clone GL1, #11-0862-85,) and anti-MHC II-PE (eBioscience, San Diego, CA, USA) clone M5/114.15.2, #12-5321-82). 7-AAD (Invitrogen, Waltham, MA, USA, #A1310) was included to stain the dead cells. Thereafter, the cells were washed with FACS buffer to remove unbound antibodies and resuspended in 100 μL FACS buffer. FACS analysis was performed on an LSR-II cytometer (BD Biosciences, Franklin Lakes, NJ, USA). The data were analyzed with FlowJo V10.

Chung C.K., Beekmann U., Kralisch D., Bierau K., Chan A., Ossendorp F, & Cruz L.J. (2022). Bacterial Cellulose as Drug Delivery System for Optimizing Release of Immune Checkpoint Blocking Antibodies. Pharmaceutics, 14(7), 1351.

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Isolation and Characterization of Primary Microglia

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Primary microglial cells were isolated from the brain via magnetic cell sorting after conjugation with anti-CD11b antibodies (Miltenyi Biotec, Bergisch Gladbach, Germany), as previously described [29 (link)]. Isolated CD11b-positive cells (>90% pure as evaluated by flow cytometry) were stained with anti-CD86-FITC (eBioscience, CA, USA), anti-CD68-APC (BioLegend, CA, USA), anti-TSPO-PE (PBR, Abcam, Cambridge, UK), and anti-CD11b-APC-Cy7 (BD Biosciences, CA, USA) antibodies after treatment with the BD Cytofix/Cytoperm Fixation/Permeabilization kit (BD Biosciences). To measure intracellular cytokines, microglia were plated in poly-d-lysine (PDL)-coated 96-well plates (BD Biosciences) and cultured in DMEM/F-12 (Gibco, CA, USA) medium supplemented with 10% fetal calf serum (Gibco) and 100 U/mL penicillium/streptomycin (Sigma-Aldrich, MO, USA) containing a leukocyte activation-cocktail with BD GolgiPlug^TM (BD Biosciences) for 12 h. Microglia cells were treated with the BD Cytofix/Cytoperm Fixation/Permeabilization kit (BD Biosciences) and then stained with anti-IL-1β-FITC (eBiosciences), anti-MIP-1α-PE (eBiosciences), anti-TNF-α-APC (BD Pharmingen, CA, USA), and anti-CD11b-APC-Cy7 (BD Biosciences) antibodies. Stained cells were analyzed using a flow cytometer (FACSCanto II; BD Biosciences).

Ano Y., Takaichi Y., Uchida K., Kondo K., Nakayama H, & Takashima A. (2018). Iso-α-Acids, the Bitter Components of Beer, Suppress Microglial Inflammation in rTg4510 Tauopathy. Molecules, 23(12), 3133.

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Evaluating CaF2:Ce,Gd,Nd NPs Impact on DCs

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In order to evaluate the effect of CaF₂: Ce, Gd, Nd NPs on immune cells, we used flow cytometry to assess the expression of DC maturation/activation markers. Briefly, murine D1 DCs and 125 µg/mL CaF₂: Ce, Gd, Nd NPs were co-cultured in a 96-well plate for 24 h in a 37 °C incubator. PBS/EDTA (Sigma-Aldrich, St. Louis, MO, USA) was used to detach the cells, which were then washed with FACS buffer and stained with anti-CD40-APC (Biolegend, San Diego, CA, USA) and anti-CD86-FITC (eBioscience, San Diego, CA, USA) antibodies. The cells were washed and resuspended in 100 μL FACS buffer after 30 min. A LSR-II cytometer (BD Biosciences, Franklin Lakes, NJ, USA) was used to measure the samples, and FlowJo (version 10) was utilized to analyze the data.

Yu Z., He Y., Schomann T., Wu K., Hao Y., Suidgeest E., Zhang H., Eich C, & Cruz L.J. (2022). Rare-Earth-Metal (Nd3+, Ce3+ and Gd3+)-Doped CaF2: Nanoparticles for Multimodal Imaging in Biomedical Applications. Pharmaceutics, 14(12), 2796.

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Modulation of BMDC Activation by AAT

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Bone marrow-derived dendritic cells (BMDC) were stimulated with IFNγ and IL-1β (5 ng/ml each, Prospec), in the absence or presence of human AAT (0.5 mg/ml). Forty-eight hours later, supernatants were collected for cytokine and nitrite analysis. In the same manner, 24 hours after stimulation, cells were examined by flow cytometry, as described (Lewis et al., 2008b (link)). The following antibodies were used for staining: anti-CD86-FITC, anti-MHC class II-PE and anti-CD11c-APC (all from eBioscience, San Diego, CA).

Baranovski B.M., Ozeri E., Shahaf G., Ochayon D.E., Schuster R., Bahar N., Kalay N., Cal P., Mizrahi M.I., Nisim O., Strauss P., Schenker E, & Lewis E.C. (2016). Exploration of α1-Antitrypsin Treatment Protocol for Islet Transplantation: Dosing Plan and Route of Administration. The Journal of Pharmacology and Experimental Therapeutics, 359(3), 482-490.

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Evaluating Macrophage Activation Markers

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The cells of each group were collected and incubated with anti-CD86-FITC (eBioscience, San Diego, CA, USA) at 4 °C in the dark for 30 min. After washing with PBS, the supernatant was removed by centrifugation; then, anti-CD206 antibody was added and incubated at 4 °C in the dark for another 30 min. The same type of antibody was used as a negative control. Finally, the expression of CD86 and CD206 in each group of cells was measured by flow cytometry and analyzed with Flowjo software (version 10).

Wen R., Luo L., Zhang R., Zhou X., Wang W, & Gong L. (2024). Structural Characterization of Polygonatum Cyrtonema Polysaccharide and Its Immunomodulatory Effects on Macrophages. Molecules, 29(9), 2076.

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Polysaccharide Extraction and Characterization from Annona squamosa

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A. squamosa was purchased from Guangzhou Tianhe fruit wholesale market, Guangzhou, China. The pulp material was identified by Professor R.M. Yu, College of Pharmacy, Jinan University, China. Standard monosaccharides and T-series dextrans were obtained from Sigma Chemical Co. (St. Louis, MO, USA). DEAE-52 cellulose and Sephadex G-100 were obtained from Whatman Ltd (Kent County, England). Sephacryl S-300 HR was obtained from Amersham Biosciences (Pharmacia, Uppsala, Sweden). The rmGM-CSF (214-14) and rmIL-4 (315-03) were obtained from PeproTechInc (Rocky Hill, NJ, USA). Anti-CD11c-FITC, anti-MHC II-PE, anti-CD86-FITC and anti-CD11c-APC antibodies were purchased from eBioscience (San Diego, CA, USA). Neutral red was purchased from Amresco (Albany, NY, USA) and the NO assay kit was supplied by the Beyotime Institute of Biotechnology (Haimen, China). Lipopolysaccharide (LPS) and FITC-dextran were purchased from Sigma–Aldrich (St. Louis, MO, USA). Anti-β-actin and anti-GAPDH antibodies were obtained from Biosharp (Beijing, China). Anti-Histone-H3 antibody was obtained from Proteintech (Wuhan, China). MAPK family antibody sampler kit and NF-κB pathway sampler kit were purchased from Cell Signaling Technology (Beverly, MA, USA). Other chemicals and reagents were of analytical grade.

Huang C., Tu W., Zhang M., Peng D., Guo Z., Huang W., Zhu J., Yu R., Song L, & Wang Y. (2023). A novel heteropolysaccharide isolated from custard apple pulp and its immunomodulatory activity in mouse macrophages and dendritic cells. Heliyon, 9(8), e18521.

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Murine Myeloid-Derived Suppressor Cell Characterization

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RPMI 1640, DMEM, FBS, and antibiotics were obtained from Life Technologies. Recombinant murine GM-CSF, anti–TGFB1-APC, and IL-2 were obtained from R&D Systems. The following antibodies were purchased from eBioscience: anti-Gr1 FITC, anti-Gr1 PE, anti-F4/80⁺ PE, anti-Cd11b⁺ PE, anti-CD11c⁺ APC, anti–PD-L1 PE, anti–PD-L1 FITC, anti–PD-L1 APC, anti–PD-L2 PE, anti-CD80 FITC, anti-CD86 FITC, anti–PD-1 APC, anti–CTLA-4 APC, anti-CD4 FITC, anti-CD8 FITC, anti–IFN-γ PE-cy7, anti-IL6 FITC, anti-IL10 APC, anti-IL12p70 PE, and functional grade anti–PD-L1 (MIH5) neutralizing antibody. For blocking, control antibody (IgG; rat IgG2b K Isotype Control Functional Grade Purified; eBioscience), anti–mouse IL-6 Functional Grade Purified neutralizing antibody (eBioscience), or anti–mouse IL-10 Functional Grade Purified neutralizing antibody (eBioscience) were used.

Noman M.Z., Desantis G., Janji B., Hasmim M., Karray S., Dessen P., Bronte V, & Chouaib S. (2014). PD-L1 is a novel direct target of HIF-1α, and its blockade under hypoxia enhanced MDSC-mediated T cell activation. The Journal of Experimental Medicine, 211(5), 781-790.

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Macrophage Polarization in Glioma Microenvironment

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GL261 cells (8 × 10⁴ − 1 × 10⁵) were seeded in the lower chambers of Transwell cell culture plate, and same amounts of RAW 264.7 cells were placed in the upper chambers at 37 °C under 5% CO₂ for 24 h in DMEM containing 10% FBS. After incubation with or without CS-I/J@CM NPs (12.5 μg/mL) for 4 h, the upper RAW 264.7 cells were removed. The GL261 cells in the lower chambers were irradiated with or without the pulsed 1064 nm laser (0.75 W/cm²) for 5 min, and the upper RAW 264.7 cells were put back and co-cultured with GL261 cells in the Transwell setup at 37 °C under 5% CO₂. After co-culturing for 24 h, the RAW 264.7 cells were collected, washed twice with PBS, and then re-dispersed in 100 μL of anti-CD86-FITC (eBioscience) and anti-CD-206-APC (eBioscience) solution and incubated for 20 min on ice. The cells were washed twice with PBS, and then analyzed with flow cytometry analysis.

Wang T., Zhang H., Qiu W., Han Y., Liu H, & Li Z. (2022). Biomimetic nanoparticles directly remodel immunosuppressive microenvironment for boosting glioblastoma immunotherapy. Bioactive Materials, 16, 418-432.

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