Mmhar 2

Manufactured by PBL Assay Science

Sourced in United States

The MMHAR-2 is a versatile laboratory instrument designed for the detection and quantification of multiple molecules in a sample. It utilizes a microfluidic-based detection system to enable high-throughput and sensitive analysis. The core function of the MMHAR-2 is to perform assays that can measure the presence and concentration of various analytes in a sample.

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Lab products found in correlation

Mmha 2, by PBL Assay Science (3 mentions) Cpg a odn2216, by InvivoGen (2 mentions) Imiquimod, by Merck Group (1 mentions) Anti ifn α, by PBL Assay Science (1 mentions) Poly 1 c, by Merck Group (1 mentions) Interleukin 2 (il 2), by Novartis (1 mentions) Igg2a, by Thermo Fisher Scientific (1 mentions) Mmhb 3, by PBL Assay Science (1 mentions) Dm irb, by Leica (1 mentions) Af1875, by R&D Systems (1 mentions) Mouse igg2a, by Thermo Fisher Scientific (1 mentions) Mouse igg1, by Thermo Fisher Scientific (1 mentions) Ab 108 c, by R&D Systems (1 mentions) Bx795, by InvivoGen (1 mentions)

6 protocols using mmhar 2

HIV Activation Regulation by IFNα/β Signaling

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Example 212

Maximal HIV Activation by Example 49 is Dependent on IFNα/β Receptor Signaling

PBMCs from HIV-infected donors on suppressive cART were isolated, as before, and treated with the compound of Example 49 (GS-9620) at the doses indicated (10 nM to 10 uM) in the presence or absence of a mouse monoclonal antibody (clone MMHAR-2) against the human Interferon α/β receptor chain 2 (PBL Assay Science, Piscataway, N.J., Cat #21385-1) used at 1:500 dilution. At day 4 after treatment, cell-free culture supernatant was collected and HIV viral RNA was quantitated using the AmliPrep/COBAS® TaqMan® assay. Fold HIV activation in PBMCs from two HIV-infected donors treated with the compound of Example 49 alone and in combination with the anti-IFNα/β mAB (IFNAR block) is depicted in FIG. 8. An 85% mean maximal reduction was seen in 4 donors (p<0.05, paired t-test).

US11116774B2. Modulators of toll-like receptors for the treatment of HIV (2021-09-14). Gilead Sciences, Inc. [US]. Inventors: Romas Geleziunas [US], Joseph E. Hesselgesser [US].

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NK Cell Activation by Dendritic Cells

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Purified NK cells were plated in a 96-well round-bottom plate (2 × 10⁶ cells/mL) and in vitro differentiated- or adult pDC were added in a NK:pDC ratio of 10:1. pDC were stimulated by adding a TLR-9 ligand (CpG-A ODN2216, 10 µg/mL, InvivoGen, San Diego, CA, USA) or a TLR-7 ligand (Imiquimod, 0.8 µg/mL, Sigma), and NK/pDC co-cultures were incubated overnight at 37 °C and 5% CO₂ atmosphere. Similarly, NK cells were co-cultured with mo-DC, peripheral blood or in vitro generated mDC. mDC and mo-DC were stimulated by adding a TLR-3 ligand (poly-IC, 10 µg/mL, Sigma). Unstimulated NK cells or NK cells cultured with unstimulated pDC were used as negative controls. NK cells stimulated with IFN-α (1000 IU/mL) were used as a positive control for each experiment. Increasing doses of IL-2 (200–20,000 IU/mL; Novartis Pharmaceuticals Canada, Dorval, Quebec, Canada) were also used to stimulate NK cells. IFN-α signaling neutralization assays were performed by incubating NK cells and pDC with neutralizing antibodies (anti-IFN-α/β receptor chain2 and anti-IFN-α) (20 µg/mL, MMHAR-2 and MMHA-2, respectively, PBL Assay Science, Piscataway, NJ, USA) for 30 min prior to the addition of TLR ligands or IFN-α. Blocking antibodies were kept in culture medium overnight.

Díaz-Rodríguez Y., Cordeiro P., Belounis A., Herblot S, & Duval M. (2017). In vitro differentiated plasmacytoid dendritic cells as a tool to induce anti-leukemia activity of natural killer cells. Cancer Immunology, Immunotherapy, 66(10), 1307-1320.

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NK Cell Activation by pDC and IFN-α

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NK cells were plated in a 96-well round-bottom plate (2×10⁶ cells/mL) and incubated overnight without stimulation or with human IFN-α (1000 UI/ml) or co-cultured with pDCs (Lin⁻HLA-DR⁺CD123⁺) (ratio NK:pDC of 10:1) and CpG-A ODN2216 (10μg/ml) (Invivogen, San Diego, CA) at 37°C in a 5% CO₂ atmosphere. Of note, pDCs and NK cells were not prepared from the same donor. In some experiments, 24-well microplates equipped with a transwell insert (0.4μm, Greiner bio-one) were used to prevent direct contact between NK cells and pDCs. For IFN-α signaling neutralization assays, NK cells were incubated for 30 minutes with neutralizing anti-IFN-α/β receptor Chain2 and anti-IFN-α antibodies (20μg/ml each) (MMHAR-2 and MMHA-2 respectively; PBL Assay Science, Piscataway, NJ), prior to the addition of IFN-α or pDCs and overnight incubation.

Lelaidier M., Dìaz-Rodriguez Y., Cordeau M., Cordeiro P., Haddad E., Herblot S, & Duval M. (2015). TRAIL-mediated killing of acute lymphoblastic leukemia by plasmacytoid dendritic cell-activated natural killer cells. Oncotarget, 6(30), 29440-29455.

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Neutralizing Autocrine and Paracrine IFN-I Effects

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Autocrine and paracrine effects of the IFN-I response were neutralized (Fig 2) by treating mDCs with cocktails of neutralizing antibodies. The anti-IFN cocktail contained antibodies neutralizing IFNAR (MMHAR-2, 5 μg/mL), IFNα (MMHA-2, 2.5 μg/mL), and IFNβ (MMHB-3, 2.5 μg/mL), all from PBL Assay Science. In the control condition, the cocktail contained corresponding control isotype antibodies, IgG1 (5 μg/mL) and IgG2a (5 μg/mL), both from ThermoFisher Scientific. One dose of antibody cocktail was administrated to the cells at the time of infection. Half a dose was added to the cell culture 1 and 3 dpi. Cells were infected at a MOI = 0.1 by wild type or mCherry-expressing MOPV or LASV. For MOPV_WT- and LASV_WT-infected mDCs, small volumes of supernatant were harvested at various times post-infection and titrated. For mCherry-expressing MOPV and LASV, mCherry fluorescence was measured by fluorescence microscopy with Leica DMIRB.

Schaeffer J., Carnec X., Reynard S., Mateo M., Picard C., Pietrosemoli N., Dillies M.A, & Baize S. (2018). Lassa virus activates myeloid dendritic cells but suppresses their ability to stimulate T cells. PLoS Pathogens, 14(11), e1007430.

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Investigating Interferon-α Modulation of CMP-001 Activity

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PBMCs were isolated, diluted to 1×10⁶ cells/mL and treated with 10 μg/mL anti-Qβ-coated CMP-001 with or without antihuman IFN-α (5 μg/mL, MMHA-2, Cat #211002; Thermo Fisher), antihuman IFNAR2 (5 μg/mL, MMHAR-2; PBL; Cat #21 385–1), mouse IgG2a, kappa isotype control (5 μg/mL, Cat# 16-4724-85; Thermo Fisher), or mouse IgG1, kappa isotype control (5 μg/mL, Cat #14471482; Thermo Fisher) for 24 hours. For CD32a neutralization experiments, monocytes were diluted to 1×10⁶ cells/mL and treated with 50 μg/mL anti-CD32a (R&D, #AF1875) or isotype control (R&D, #AB-108-C) for 15 min followed by treatment with 10 μg/mL anti-Qβ-coated CMP-001 and 1.792×10⁴ U/mL recombinant IFN-α for 18–20 hours.

Sabree S.A., Voigt A.P., Blackwell S.E., Vishwakarma A., Chimenti M.S., Salem A.K, & Weiner G.J. (2021). Direct and indirect immune effects of CMP-001, a virus-like particle containing a TLR9 agonist. Journal for Immunotherapy of Cancer, 9(6), e002484.

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Inhibition of Interferon Signaling in HMPV

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Human recIFN-β and -λ1 were purchased from Peprotech. NIFNλR1 (MMHLR-1) and nIFNAR2 (MMHAR-2; interferon-α/β receptor 2) were purchased from PBL. Cells were incubated with nIFNλR1 (10 µg/mL) or nIFNAR (10 µg/mL) 30 min before infection with virus. RecIFN-β (1000 U/mL) or recIFN- λ1 (1 µg/mL) were added 15 h after infection with virus for a total of 3 h. Cells were incubated with the pharmacological TBK1 inhibitor BX795 (10 µM; InvivoGen) 30 min prior to infection with HMPV if not indicated otherwise. Cells were incubated with either 5 µM or 10 µM S-Ruxolitinib 2 h prior to infection with virus.

Loevenich S., Spahn A.S., Rian K., Boyartchuk V, & Anthonsen M.W. (2021). Human Metapneumovirus Induces IRF1 via TANK-Binding Kinase 1 and Type I IFN. Frontiers in Immunology, 12, 563336.

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