Cd83 apc hb15e

Manufactured by BioLegend

CD83-APC (HB15e) is a fluorescently labeled antibody that binds to the CD83 antigen. CD83 is a cell surface protein expressed on mature dendritic cells and other antigen-presenting cells. This product is intended for research use only.

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Lab products found in correlation

Cd3 apc, by BioLegend (1 mentions) Streptavidin fitc, by Agilent Technologies (1 mentions) Cd86 apc, by BioLegend (1 mentions) Cd11b pecy7 m1 70, by BioLegend (1 mentions) Ly6c biotin al 21, by BD (1 mentions) Cd45r b220 percp cy5, by Thermo Fisher Scientific (1 mentions) Cd19 pc5, by Beckman Coulter (1 mentions) Ly6g pe, by BD (1 mentions) Lsrfortessa, by BD (1 mentions) Cd14 qdot605 clone tuk4, by Thermo Fisher Scientific (1 mentions) Cd66a c e asl 32, by BioLegend (1 mentions) Cd40 percp cy5.5 5c3, by BioLegend (1 mentions) Cd86 pe it2, by BD (1 mentions) Kc57 rd1, by Beckman Coulter (1 mentions) Hla dr percp, by BD (1 mentions) Perm wash, by BD (1 mentions)

3 protocols using cd83 apc hb15e

Surface Staining of Immune Cells

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For surface staining of human specimens, cell suspensions were incubated with the appropriate combination of the following monoclonal antibodies: CD19-PC5 (J3-119, Beckman Coulter), CD80-Brilliant Violet 421™ (2D10, BioLegend^®), CD83-APC (HB15e, BioLegend^{^®}), CD86-APC (IT2.2, BioLegend^®), and HLA-DR-V500 (G46-6, BD Horizon™). For surface staining of mouse specimens, cell suspensions were incubated with Fc-blocking antibody (Bioxcell, 2.4G2) to avoid unspecific Fc-Receptor binding. After washing, the cells were incubated with the appropriate combination of the following antibodies: CD11b-PECy7 (M1/70, BioLegend^®), Ly6G-PE (1A8, BD Biosciences), Ly6C-Biotin (AL-21, BD Biosciences), CD3-APC(17A2, BioLegend^®), CD45R-B220-PerCP-Cy5.5 (RA3-6B2, eBiosciences). To detect anti-Ly6C-Biotin antibody binding, cells were subsequently stained with streptavidin-FITC (Dako). The samples were acquired with BD LSRFortessa (BD Biosciences), and results were analyzed with FlowJo software (Tree Star, Inc.).

Ferreira T.P., Mariano L.L., Ghilosso-Bortolini R., de Arantes A.C., Fernandes A.J., Berni M., Cecchinato V., Uguccioni M., Maj R., Barberis A., Silva P.M, & Martins M.A. (2016). Potential of PEGylated Toll-Like Receptor 7 Ligands for Controlling Inflammation and Functional Changes in Mouse Models of Asthma and Silicosis. Frontiers in Immunology, 7, 95.

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Whole Blood and PBMC Immune Activation Protocol

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Ultrapure lipopolysacharide (LPS, TLR4 ligand, 100ng/mL used in whole blood assays) isolated from Salmonella minnesota, was obtained from InvivoGen. PAM3 (Pam3CSKKKK, N-palmitoyl-S-[2,3-bis- (palmitoyloxy)-propyl]-(R)-cysteinyl-(lysyl)3-lysine, TLR2/1 ligand, 300ng/mL used in whole blood assays, 10 µg/mL used in PBMC phosphoflow assays) was obtained from EMC Microcollections. Log-phase derived live BCG expressing green fluorescent protein (BCG-GFP, Pasteur strain; used at 3.5 × 10⁵ CFU/mL in assays) was donated by Muazzam Jacobs, University of Cape Town.
The following antibodies were used for whole blood flow cytometry: CD14-QDot605 (clone Tuk4, Invitrogen), TNF-α-PECy7 (Mab11), CD11c-V450 (B-ly6), CD86 (B70/B7–2)-PE (both from BD Pharmingen), CD11c-PerCPCy5.5 (Bu15), HLA-DR-AlexaFluor700 (L243), IL-12/23p40-Pacific Blue (C11.5), IL-10-PE (JES3-19F1), IL-6-APC (MQ2-13A5), CD40-PerCP-Cy5.5 (5C3) and CD83-APC (HB15e) (all from Biolegend). CD66a/c/e (ASL-32, Biolegend) was conjugated in-house to QDot565 (Invitrogen) using the manufacturer’s protocol. The following antibodies were used for PBMC phosphoflow experiments: CD14-Pacific Blue (clone M5E2), p-p38 MAPK-PECy7 (36/p38: pT180/pY182), p-Erk1/2-AlexaFluor647 (20A: pT202/pY204), IκBα-PE (25/IκBα/MAD-3), all from BD Biosciences.

Shey M.S., Nemes E., Whatney W., de Kock M., Africa H., Barnard C., van Rooyen M., Stone L., Riou C., Kollmann T., Hawn T.R., Scriba T.J, & Hanekom W.A. (2014). Maturation of Innate Responses to Mycobacteria over the First 9 Months of Life. Journal of immunology (Baltimore, Md. : 1950), 192(10), 4833-4843.

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Multiparameter Immunophenotyping of T-Cells

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To determine CD86, CD83 and HLA-DR on DCs the cells were washed twice using FACS buffer and were subsequently stained using CD86-PE (IT2.2, BD Biosciences), CD83-APC (HB15e, Biolegend) and HLA-DR-PerCp (L243, BD Biosciences) for 30min at 4°C in FACS buffer. Cells were washed once more with FACS buffer and were analysed on a FACS Canto II apparatus. To assess the capacity of T-cells to produce cytokines/ chemokines, cells were permeabilized with PermWash (BD Pharmingen) according to manufacturer’s protocol to stain for intracellular markers. Cells were stained for p24-RD1 (Beckman Coulter, KC57-RD1) and IFN-γ-FitC (4S.B3), IL2-PerCp-Cy5.5 (MQ1-17H12), IL-4-APC (MP4-25D2), TNF-α-PE-CF594 (MAb11), Mip-1β-AlexaFluor700 (D21-1351) (all from BD Bioscience) for 30min at 4°C in PermWash. Next, these cells were washed once with PermWash, resuspended in FACS buffer (PBS+ 2%FCS) and measured on a FACS Canto II apparatus. The histograms depict all T-cells capable of producing a certain cytokine/ chemokine at the optimal time point. The optimal time point was determined by the percentage of live cells (>50%) and the level of HIV-1 infection, which varied per donor and per virus between day 5 and 7.

Mouser E.E., Pollakis G., Yazdanbakhsh M., Harnett W., de Jong E.C, & Paxton W.A. (2016). Brugia malayi Antigen (BmA) Inhibits HIV-1 Trans-Infection but Neither BmA nor ES-62 Alter HIV-1 Infectivity of DC Induced CD4+ Th-Cells. PLoS ONE, 11(1), e0146527.

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