Anti cd86 pe cy5

Manufactured by BD

Sourced in United States

Anti-CD86-PE-Cy5 is a fluorescently labeled monoclonal antibody that binds to the CD86 protein. CD86 is a costimulatory molecule expressed on the surface of antigen-presenting cells. The PE-Cy5 conjugate allows for the detection and analysis of CD86-expressing cells using flow cytometry.

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Lab products found in correlation

Anti cd14 pe, by BD (1 mentions) Anti hla dr fitc, by BD (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) T75 flask, by Corning (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Gm csf, by Thermo Fisher Scientific (1 mentions) Macs cd14 microbeads, by Miltenyi Biotec (1 mentions) Anti cd3 pb, by BioLegend (1 mentions) Anti cd163 alexa fluor 647, by BioLegend (1 mentions) Anti cd80 pe cy7, by BioLegend (1 mentions) Recombinant mouse il 4, by Thermo Fisher Scientific (1 mentions) Anti il 4 pe, by BD (1 mentions) Anti cd4 v450, by BD (1 mentions) Anti cd40 apc, by BD (1 mentions) Anti xbp1, by Cell Signaling Technology (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) Anti rabbit igg fitc, by Thermo Fisher Scientific (1 mentions) Anti tnf α pe, by BD (1 mentions) Anti pdl 1 biotin, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

CD86-PE (81 citations) Anti-CD86 (74 citations) Anti-CD86-PE (43 citations)

5 protocols using anti cd86 pe cy5

Monocyte-derived Dendritic Cell Activation

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PBMC were thawed and monocytes were isolated by positive selection using MACS CD14 MicroBeads (MiltenyiBiotec, Auburn, CA, USA) following manufacturer’s instructions. Monocytes were seeded in T75 flasks (Corning) in RPMI medium containing 10% heat-inactivated fetal calf serum (FCS, 50 U/ml penicillin, 50 µg/ml streptomycin (Gibco, Paisley, United Kingdom), IL-4 and GM-CSF (Biosource International, Camarillo, CA, USA) both at 10 ng/ml for 6 days to generate monocyte-derived immature DCs. Mature DCs were generated by stimulating DCs with 10 ng/ml lipopolysaccharide (LPS) for an additional 24 hours. DC differentiation and maturation was validated by flow cytometry using anti-CD14 PE (BD Pharmingen), anti-HLA-DR FITC (BD Pharmingen), anti-CD1a Alexa Fluor 700 (Biolegend), anti-CD163 Alexa Fluor 647 (Biolegend), anti-CD80 PE-Cy7 (Biolegend), anti-CD86 PE-Cy5 (BD Pharmingen) and anti-CD3 PB (Biolegend).
Matured DCs were loaded with Rv2034 protein, Rv2034 p81–100 and different conditions of Mtb lysate and incubated for 24 hours. Cells were washed and T-cell clone added. After 2 hours BFA was added and culture incubated o/n. Activation of T cells was determined by detection of CD154 and Th1 markers using the T-helper subset panel.

Commandeur S., Coppola M., Dijkman K., Friggen A.H., van Meijgaarden K.E., van den Eeden S.J., Wilson L., van der Ploeg-van Schip J.J., Franken K.L., Geluk A, & Ottenhoff T.H. (2014). Clonal Analysis of the T-Cell Response to In Vivo Expressed Mycobacterium tuberculosis Protein Rv2034, Using a CD154 Expression Based T-Cell Cloning Method. PLoS ONE, 9(6), e99203.

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Comprehensive Murine Immune Cell Analysis

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Antibodies were purchased as followed: Anti-CD4-V450, anti-CD8α-APCcy7, anti-B220-V450, anti-CD138-Pe-cy7, anti-CD86-Pe-cy5, anti-FAS-PE, anti-GL-7-APC, anti-CD40-APC, anti-MHCII-FITC, anti-IL-4-PE, anti-IL-5-PE, anti-IgM-Pe-cy7, anti-IgG1-APC, anti-IFN-r-Percp5.5, anti-TNF-α-PE, and anti-IL-17-Pe-cy7 were purchased from BD Biosciences (Franklin Lakes, NJ). Anti-PDL-1-biotin (eBioscience, San Diego, CA), anti-XBP-1s (Cell Signaling Technology, Danvers, MA), and anti-Rabbit IgG-FITC (Thermo Fisher Scientific, Waltham, MA) antibodies were purchased from commercial sources. Recombinant mouse IL-4 (PeproTech, Rocky Hill, NJ) and LPS (Sigma-Aldrich, St. Louis, MO) were purchased from the commercial companies. Goat F(ab’)2 Anti-Mouse IgM (1022-01, SouthernBiotech, Birmingham, AL) and anti-mouse CD40 (BE0016-2, BioXCell, Lebanon, NH) were commercially purchased from companies.

Choi H.J., Tang C.H., Tian L., Wu Y., Sofi M.H., Ticer T., Schutt S.D., Hu C.C, & Yu X.Z. (2021). XBP-1s Promotes B Cell Pathogenicity in Chronic GVHD by Restraining the Activity of Regulated IRE-1α-Dependent Decay. Frontiers in Immunology, 12, 705484.

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Vaccine-Induced Dendritic Cell Activation

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DC activation was tested by removing sciatic and popliteal lymph nodes 24 hours and 48 hours after vaccination with different vaccine formulations, as described earlier. The lymph nodes were harvested and prepared into a cell suspension. The lymph node cells were then washed, blocked, and stained with antibodies (anti-CD11c-FITC, anti-major histocompatibility complex class II [MHCII]-PE and anti-CD86-PE-Cy5; all from BD Biosciences), and flow cytometry analysis was performed on a BD FACSVerse™ flow cytometer.

Liu Z., Luo L., Zheng S., Niu Y., Bo R., Huang Y., Xing J., Li Z, & Wang D. (2016). Cubosome nanoparticles potentiate immune properties of immunostimulants. International Journal of Nanomedicine, 11, 3571-3583.

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DC Maturation and Phagocytosis Assays

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Immature DCs were cultured alone or in co-culture with AMO-1 pre-treated for 48 h in the presence of lipopolysaccharide (LPS) or DMSO or SIX2G 60 nM, at a 5:1 Effector: Target (E:T) ratio. After 24 h of co-culture, cells were collected, washed in PBS and then stained with anti–CD86-PeCy5 (#555666, BD, Franklin Lakes, NJ, USA), and 7AAD (#51-68981E, BD, Franklin Lakes, NJ, USA). Dead cells were excluded by 7AAD positivity, and CD86 DC maturation marker expression was evaluated by flow cytometry.
For phagocytosis assay, AMO-1 were labelled with CellTrace Far Red (Thermo Fisher Scientific, Waltham, MA, USA), treated with DMSO or SIX2G 60 nM for 48 h, and then co-cultured for 12 h at a 5:1 ratio with monocyte-derived DCs (Mo-DCs). Cells were then detached, collected and stained with CD11b-PeCy7 (#557743, BD, Franklin Lakes, NJ, USA) and 7AAD viable marker (BD, Franklin Lakes, NJ, USA) to exclude apoptotic cells. A phagocytosis analysis was performed by flow cytometry. The percentage of phagocytosed cells was evaluated as the percentage of 7AAD⁻ CD11b⁺ Far Red⁺ cells.

Grillone K., Riillo C., Rocca R., Ascrizzi S., Spanò V., Scionti F., Polerà N., Maruca A., Barreca M., Juli G., Arbitrio M., Di Martino M.T., Caracciolo D., Tagliaferri P., Alcaro S., Montalbano A., Barraja P, & Tassone P. (2022). The New Microtubule-Targeting Agent SIX2G Induces Immunogenic Cell Death in Multiple Myeloma. International Journal of Molecular Sciences, 23(18), 10222.

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Comprehensive Flow Cytometry Analysis of PBMC

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Flow cytometry analysis was performed prospectively using samples from a part of patients at treatment initiation and 6 months post-treatment. Peripheral blood mononuclear cells were separated using density gradient centrifugation with the Ficoll-Paque Plus (GE Healthcare, Uppsala, Sweden) and cryopreserved in the CELLBANKER 1 (Nippon Zenyaku Kogyo, Fukushima, Japan). The antibodies used for staining peripheral blood mononuclear cells were as follows; anti-CD4-VioGreen (Miltenyi Biotec, Bergisch Gladbach, Germany); anti-CD3-Paci c Blue/ uorescein isothiocyanate (FITC), anti-CD8-Paci c Blue, anti-CD14-(APC)-Cy7, anti-CD20 allophycocyanin-cyanine 7 (APC-Cy7), anti-CD25 phycoerythrin (PE)-Cy5, anti-CD27-PE-Cy7, anti-CD38-PE-Cy5, anti-CD45RO-PE-Cy7, anti-CD56-PE/PE-Cy7, anti-CD80-FITC, anti-CD86-PE-Cy5, anti-CD127-FITC, anti-CD161-APC and anti-chemokine (C-X-C motif) receptor 3 (CXCR3)-PE (all from BD Biosciences, Franklin Lakes, NJ, USA); anti-CD16-Brilliant Violet 510 and anti-CCR6-Brilliant Violet 421 (both from BioLegend, San Diego, CA, USA); and anti-mouse immunoglobulin G isotypematched controls (VioGreen from Miltenyi Biotec, the others from BD Biosciences). The peripheral cell subsets identi ed have been described in Supplementary table 1.

Inamo J., Kaneko Y., Kikuchi J, & Takeuchi T. (2021). High serum IgA and activated Th17 and Treg predict the efficacy of abatacept in patients with early, seropositive rheumatoid arthritis. Clinical rheumatology, 40(9).

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