Anti ifnar2

Manufactured by PBL Assay Science

Sourced in United States

Anti-IFNAR2 is a monoclonal antibody that specifically binds to the IFNAR2 subunit of the type I interferon receptor. It is used as a tool in research applications to study the function and signaling of type I interferons.

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Lab products found in correlation

Tofacitinib, by Apexbio (1 mentions) Tocilizumab, by Selleck Chemicals (1 mentions) Anti il 10, by BioLegend (1 mentions) Anti il 10r, by BioLegend (1 mentions) Anti ifn β, by R&D Systems (1 mentions) Cpg2006, by InvivoGen (1 mentions) Cd3 bv510, by BioLegend (1 mentions) Cd56 bv711, by BioLegend (1 mentions) Cd19 af700, by BioLegend (1 mentions) Cd14 af488, by BioLegend (1 mentions) Lsrfortessa, by BD (1 mentions)

3 protocols using anti ifnar2

Whole Blood Cytokine Modulation Assay

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Whole blood was incubated for 4 h with tofacitinib (500 nM, ApexBio) or tocilizumab (50 μg ml⁻¹, Selleckchem) or vehicle controls (DMSO and human IgG control, 50 μg ml⁻¹, BioLegend, respectively). For IFN-I blocking, whole blood was incubated with a combination of anti-IFNAR2 (2.5 μg ml⁻¹ PBL Assay Science), anti-IFNα (0.2 μg ml⁻¹, PBL 31110–1) and IFNβ (0.2 μg ml⁻¹, PBL 31401-1) antibodies or vehicle control for 4 h. For IL-10 blocking, whole blood was incubated with a combination of anti-IL-10 (5 μg ml⁻¹, BioLegend) and anti-IL-10R (5 μg ml⁻¹, BioLegend) for 4 h. After blocking, whole blood was fixed using Proteomic Stabilizer PROT1 (1.4× blood volume, SmartTube) for 10 min at room temperature and frozen at −80 °C.

Malle L., Patel R.S., Martin-Fernandez M., Stewart O.J., Philippot Q., Buta S., Richardson A., Barcessat V., Taft J., Bastard P., Samuels J., Mircher C., Rebillat A.S., Maillebouis L., Vilaire-Meunier M., Tuballes K., Rosenberg B.R., Trachtman R., Casanova J.L., Notarangelo L.D., Gnjatic S., Bush D, & Bogunovic D. (2023). Autoimmunity in Down’s syndrome via cytokines, CD4 T cells and CD11c+ B cells. Nature, 615(7951), 305-314.

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CpG Oligodeoxynucleotide Signaling in IFN-β Response

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Synthetic phosphodiester oligodeoxynucleotides (CpG 2006 and GpC 2006) were synthesized by InvivoGen (Toulouse, France) and used at indicated concentration. IFN-β was used at 100 or 1000 UI/ml (Avonex, Biogen, Nanterre Cedex, France), anti-IL-29, anti-IFNβ (R&D) and anti-IFNAR2 (PBL, New Jersey, NJ, USA). Cells were stimulated overnight and the response was monitored by luciferase assay or enzyme-linked immunosorbent assay. For luciferase assay transient transfection of the reporter plasmid NF-kB, or ELAM luciferase was performed as previously described.^{41 (link)} Enzyme-linked immunosorbent assays were done in accordance to manufacturer's instructions (R&D system).

Parroche P., Roblot G., Le Calvez-Kelm F., Tout I., Marotel M., Malfroy M., Durand G., McKay J., Ainouze M., Carreira C., Allatif O., Traverse-Glehen A., Mendiola M., Pozo-Kreilinger J.J., Caux C., Tommasino M., Goutagny N, & Hasan U.A. (2016). TLR9 re-expression in cancer cells extends the S-phase and stabilizes p16INK4a protein expression. Oncogenesis, 5(7), e244-.

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Profiling Interferon Signaling in Down Syndrome

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Phospho-STAT1 staining: Whole blood from individuals with DS and healthy controls were subject to Ficoll gradient to collect the mononuclear cell layer (PBMC) and frozen. For direct stimulation, thawed PBMCs were incubated in complete RPMI overnight, washed, stained with live-dead in PBS for 10 min at 37°C, and finally were stimulated for 15 min with indicated amounts of IFNα2b. For staining, cells were washed 2 times with 0.5% BSA and surface-stained on ice for 30 minutes (CD16-AF647, CD14-AF488, CD3-BV510, CD19-AF700, CD56-BV711, all from Biolegend) followed by fixation/permeabilization in 90% ice-cold methanol and staining with PE anti-phospho-STAT1 Y701 (1:25, BD).
IFNAR1 and IFNAR2 staining: hTERT-immortalized fibroblasts were scraped off in 5mM EDTA, washed, and stained with live-dead in PBS for 30 min on ice. They were then washed and stained with anti-IFNAR1 (clone AA3, courtesy of Sandra Pellegrini in Supplemental Fig 1A and clone MARI-5A3 from Millipore Sigma for Supplemental Fig 3A) or anti-IFNAR2 (PBL) for 2 hours on ice. The cells were washed and stained with biotin-conjugated rat anti-mouse IgG (H+L) (Thermo Fisher) for 40 min on ice, followed by PE-conjugated Streptavidin for 10 min on ice.
Flow cytometry was acquired on a Cytek Aurora or BD LSR Fortessa, and data were analyzed with Cytobank (https://www.cytobank.org/).

Malle L., Martin-Fernandez M., Buta S., Richardson A., Bush D, & Bogunovic D. (2022). Excessive Negative Regulation of Type I Interferon Disrupts Viral Control in Individuals with Down Syndrome. Immunity, 55(11), 2074-2084.e5.

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