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Anti cd40 apc

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Anti-CD40-APC is a monoclonal antibody that binds to the CD40 antigen. It is conjugated to the fluorescent dye Allophycocyanin (APC).

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Lab products found in correlation

Anti cd86 fitc, by Thermo Fisher Scientific (2 mentions) Lsrii cytometer, by BD (2 mentions) Anti mhc 2 pe, by Thermo Fisher Scientific (1 mentions) 7 aad, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Cd86 fitc, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Lipopolysaccharide, by Thermo Fisher Scientific (1 mentions) Aqueous mts reagent powder, by Promega (1 mentions) Phalloidin ifluor 488 reagent, by Abcam (1 mentions) Pbs edta, by Merck Group (1 mentions) Version 10, by FlowJo (1 mentions) Flowlogic software, by Miltenyi Biotec (1 mentions) Anti cd86 bv421, by BD (1 mentions) Anti hla abc fitc, by BD (1 mentions) Anti cd11c alexafluor700, by BD (1 mentions) Anti cd83 pe, by BD (1 mentions) Anti cd4 apc cy7, by BioLegend (1 mentions) Anti cd8 pe, by BioLegend (1 mentions) Anti pd l1 apc, by BioLegend (1 mentions)

Spelling variants of this product

Anti-CD40 (36 citations) CD40-APC (18 citations) Anti-mouse CD40-APC (4 citations) APC-anti-CD40 (clone 3/23, (2 citations) Anti-human CD40-APC (2 citations) APC-labeled anti-mouse CD40 (2 citations)

8 protocols using anti cd40 apc

1

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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2

Characterization of Lanthanide-Treated Immune Cells

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The following chemicals were acquired from Sigma-Aldrich (St. Louis, MO, USA): neodymium (III) chloride hexahydrate (NdCl3·6H2O, 99.9%), calcium chloride dihydrate (CaCl2·2H2O, 99.5%), cerium (III) chloride heptahydrate (CeCl3·7H2O, 99.9%), potassium citrate tribasic monohydrate (HOC(COOK)(CH2COOK)2·H2O, ≥99.0%), gadolinium (III) chloride hexahydrate (GdCl3·6H2O, 99.9%), ammonium fluoride (NH4F, ≥98.0%). Biolegend (San Diego, CA, USA) supplied the anti-CD40-APC. Thermo Fisher Scientific (Waltham, MA, USA) provided fetal calf serum (FCS), 4′,6-Diamidino-2-Phenylindole (DAPI), dulbecco’s Modified Eagle’s Medium (DMEM) and CD86-FITC. Promega (Madison, WI, USA) offered cell titer 96 AQueous MTS Reagent Powder. PeproTech (Cranbury, NJ, USA) supplied lipopolysaccharide (LPS). Bioline (London, UK) delivered agarose. Abcam (Cambridge, UK) supplied the phalloidin- iFluor 488 Reagent. All of water used in the experiments was ultrapure deionized water.
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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3

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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4

Activated myDCs response to T-VEC

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Upon treatment, BDCA-1+ myDCs were seeded in a 96-well round bottom plate at a density of 150,000–200,000 cells per well. BDCA-1+ myDCs were treated with either active T-VEC (MOI 10), heat-inactivated T-VEC (MOI 10; heat-inactivation: 15 min 65 °C, 1 min 100 °C) or conditioned medium, i.e., SN of dying 624-mel and 938-mel 24 h and 48 h after treatment with T-VEC (MOI 1). Untreated BDCA-1+ myDCs served as a negative control. As a positive control, BDCA-1+ myDCs were treated with a mix containing ssRNA fragments derived from HIV-1 long terminal repeat and protamine sulfate (PS/LTR). After overnight incubation, cells were harvested and stained with anti-CD11c-AlexaFluor 700 (BD Biosciences, 561352), anti-CD1c-BV510 (BD Biosciences, 742747), anti-CD80-PerCP-eFluor710 (ThermoFisher Scientific, 46-0809-42), anti-CD86-BV421 (BD Biosciences, 562432), anti-CD40-APC (BioLegend, 334310), anti-CD83-PE (BD Biosciences, 556855), anti-CD274-PE-CF594 (BD Biosciences, 563742) and anti-HLA-ABC-FITC (BD Biosciences, 557348) for 20 min at 4 °C and in the dark. Cells were acquired on the flow cytometer (BD LRS Fortessa) and data were analyzed with with FlowLogic software (Miltenyi Biotec, Version 7.3).
Kalus P., De Munck J., Vanbellingen S., Carreer L., Laeremans T., Broos K., Dufait I., Schwarze J.K., Van Riet I., Neyns B., Breckpot K, & Aerts J.L. (2022). Oncolytic Herpes Simplex Virus Type 1 Induces Immunogenic Cell Death Resulting in Maturation of BDCA-1+ Myeloid Dendritic Cells. International Journal of Molecular Sciences, 23(9), 4865.
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5

Comprehensive Immune Profiling of Pancreatic Tumors

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Pancreatic tumour cell lines, MDDC and T-cells were assessed for expression of different markers using the following antibodies: Anti-PDL-1 APC, anti-Galectin 9 PE, anti-CD39 FITC, anti-CD47 Percp-Cy5.5, anti-HLA-class I Alexa Flour 700, anti-MIC A/B PE, anti-ULBP1 FITC, anti-ULBP 2,5 6 Percp, anti-ULBP 3 APC,anti-CD86 PE, anti-CCR7 PE-CY7, anti-CD40 APC, anti-HLA-class II PE-CY5 anti-IFN-γ PE-CY7, anti-CD3 Alexa Flour 488, anti-CD4 APC-CY7, anti-CD8-PE and anti-CD69 APC (Biolegend, CA).
Smith P.L., Yogaratnam Y., Samad M., Kasow S, & Dalgleish A.G. (2020). Effect of Gemcitabine based chemotherapy on the immunogenicity of pancreatic tumour cells and T-cells. Clinical & Translational Oncology, 23(1), 110-121.
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6

Murine Dendritic Cell Activation Assay

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Cells were isolated from the bone marrow of C57BL/6J mice and were cultured in RPMI 1640 medium supplemented with 20 ng/mL GM-CSF and 10 ng/mL IL-4. The culture medium was replaced and the cytokines were replenished every two days. Nonadherent and loosely adherent cells were harvested on Day 8 and plated in 24-well plates. The cells (1×106 cells/mL/well) were cultured with compounds (0.1% DMSO served as a control) for 24 h at 37 °C and stained with fluorescein-conjugated anti-CD11c-PerCP/Cy5.5 (cat. no. 117328; BioLegend, San Diego, USA), anti-CD40-APC (cat. no. 124612; BioLegend), anti-CD80-FITC (cat. no. 104706; BioLegend) and anti-CD86-BV650 (cat. no. 105036; BioLegend).
Ren S., Wang X, & Jin G. (2022). Conjugate of ibrutinib with a TLR7 agonist suppresses melanoma progression and enhances antitumor immunity. International Journal of Biological Sciences, 18(1), 166-179.
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7

Immune Cell Profiling in Immunized Mice

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Quadriceps muscles from immunized mice were harvested after 18–24 h of the injection and cut into small pieces. A single cell suspension was prepared using skeletal muscle dissociation kit (Miltenyi Biotec) according to manufacturer’s protocol. Briefly, small pieces of muscle were incubated in enzyme solution (enzyme D, enzyme P and enzyme A in DMEM) for 30 min at 37oC with gentle shaking, passed through 70 µm cell strainer and wash with 10 ml DMEM. Centrifuge cell suspension at 300xg for 20 min. Cells were stained with a combination of the following fluorescently labeled antibodies: anti-CD45-AF700, anti-CD3-BV786, anti-CD19-BV605, anti-CD49b-PE, anti-CD11b-PE-Cy5, anti-CD11c-BV711, anti-F4/80-FITC, anti-Ly6C-PerCp, anti-Ly6G-PE/Cy7, anti-MHCII-PE/Dazzle594, anti-CD80-BV650, anti-CD40-APC, anti-CXCR3-BV510 and anti-CCR2-BV421 (all from BioLegend). Live/dead cells were analyzed using fixable viability dye efluor780 (eBioscience). The stained cells were acquired using BD LSRFortessa cell analyzer and data was analyzed using FlowJo software (Tree Star). Total immune cells (CD45+), myeloid cells (CD11b+), monocytes (CD11b+Ly6C+), macrophages (CD11b+F4/80+), neutrophils (CD11b+Ly6G+), dendritic cells (DCs; CD11c+MHCII+), natural killer cells (NK cells; CD49b+, CD3−), B cells (CD11b−CD11c−CD19+), T cells (CD11b−CD11c−CD3+) and NKT cells (CD49b+CD3+) were gated by flow cytometry.
Jain S., George P.J., Deng W., Koussa J., Parkhouse K., Hensley S.E., Jiang J., Lu J., Liu Z., Wei J., Zhan B., Bottazzi M.E., Shen H, & Lustigman S. (2018). The parasite-derived rOv-ASP-1 is an effective antigen-sparing CD4+ T cell-dependent adjuvant for the trivalent inactivated influenza vaccine, and functions in the absence of MyD88 pathway. Vaccine, 36(25), 3650-3665.
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8

Multicolor Flow Cytometry Panel

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Anti‐B220 Cy5, anti‐B220 Cy3, anti‐CD62L Pacific Orange, anti‐CD4 Alexa488, anti‐IgD Cy5 and anti‐Lyve‐1 Cy3 were provided by E. Kremmer (Helmholtz Zentrum München, Munich, Germany). Anti‐CD4 PerCp, anti‐CD40 APC, anti‐CD80 APC, and anti‐CD45.1 PE were purchased from Bio‐legend. Anti‐CD11b PE‐Cy7, Anti‐CD11c PE‐Cy7, anti‐CD3 PE, anti‐B220 PE, and anti‐CD45.1 FITC were purchased from eBioscience, San Diego, CA. Anti‐CD11b PE was purchased from Caltag Laboratories, Anti‐CD11c PE from BD Biosciences, and anti‐pDCA‐1 APC from Milteyni Biotec.
Kohli K., Janssen A, & Förster R. (2016). Plasmacytoid dendritic cells induce tolerance predominantly by cargoing antigen to lymph nodes. European Journal of Immunology, 46(11), 2659-2668.
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