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Anti mouse ifnar1 clone mar1 5a3

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Anti-mouse IFNAR1 (Clone: MAR1-5A3) is a monoclonal antibody that binds to the mouse interferon-alpha/beta receptor 1 (IFNAR1) subunit. This antibody is commonly used in research applications to block or neutralize the type I interferon signaling pathway in mouse models.

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Lab products found in correlation

Adu s100, by Chemietek (1 mentions) Bortezomib, by Selleck Chemicals (1 mentions) Anti mouse cd16 cd32, by Thermo Fisher Scientific (1 mentions) Goat anti mouse igg h l alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Macsquant analyzer 10, by Miltenyi Biotec (1 mentions) Cellstripper, by Corning (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Rat igg2a isotype control, by BioXCell (1 mentions) Mouse igg1 isotype control, by BioXCell (1 mentions) Lookout mycoplasma pcr detection kit, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)

4 protocols using anti mouse ifnar1 clone mar1 5a3

1

Bortezomib, IFNAR1, and ADUS-100 Assay

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Bortezomib was purchased from Selleckchem (S1013) and resuspended in DMSO. Anti-mouse IFNAR1 (Clone: MAR1-5A3) (#BE0241) MoAb and mouse IgG1 Isotype CNT MoAb were purchased from BioXCell and resuspended in InVivoPure Diluition Buffer following manufacturer’s instructions. ADUS-100 (#CT-ADUS100) was purchased from ChemieTek.
Gulla A., Morelli E., Samur M.K., Botta C., Hideshima T., Bianchi G., Fulciniti M., Malvestiti S., Prabhala R.H., Talluri S., Wen K., Tai Y.T., Richardson P.G., Chauhan D., Sewastianik T., Carrasco R.D., Munshi N.C, & Anderson K.C. (2021). Bortezomib Induces Anti–Multiple Myeloma Immune Response Mediated by cGAS/STING Pathway Activation. Blood Cancer Discovery, 2(5), 468-483.
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2

Cell Surface Antigen Staining Protocol

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For staining of cell surface antigen, cells were detached with Cellstripper (Corning) and resuspended in PBS with 0.5% FBS and 1 mM EDTA (FACS [fluorescence-activated cell sorting] buffer). Bst2 was stained with phycoerythrin (PE)-labeled anti-Bst2 (BioLegend, clone 129C1; 1:500 in FACS buffer) and anti-mouse CD16/CD32 (eBioscience; 1:250 in FACS buffer) on ice for 45 min. Ifnar1 was stained with 1 μg/ml of anti-mouse Ifnar1 (clone MAR1-5A3; BioXcell) at room temperature for 45 min, followed by incubation with a goat anti-mouse IgG (H+L) Alexa Fluor 647 (Thermo Fisher, catalog no. A21236; 1:1,000 in FACS buffer) at room temperature for 30 min. For cells infected with GFP-labeled viruses (SINV-EEEV-GFP and VSV-GFP), cells were detached with trypsin and fixed with 1% paraformaldehyde (PFA) at room temperature for 15 min. Flow cytometry was performed on a MACSQuant Analyzer 10 (Miltenyi Biotec), and data were analyzed using FlowJo 10.6.1.
Ma H., Qian W., Bambouskova M., Collins P.L., Porter S.I., Byrum A.K., Zhang R., Artyomov M., Oltz E.M., Mosammaparast N., Miner J.J, & Diamond M.S. (2020). Barrier-to-Autointegration Factor 1 Protects against a Basal cGAS-STING Response. mBio, 11(2), e00136-20.
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3

Murine Cancer Cell Lines and Reagents

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BALB/c-derived mouse TSA mammary carcinoma cell line was obtained from Dr. Lollini (19 (link)) and authenticated by IDEXX Bioresearch (Columbia, MO, USA) in 2019. C57BL/6-derived mouse MCA38 colorectal carcinoma cells line was obtained from Dr. Frey (20 (link)) in 2009 and authenticated by IDEXX Bioresearch in 2016. Both cells were grown in complete medium consisting of DMEM (Invitrogen) supplemented with L-glutamine (2 mol/L), penicillin (100 U/mL), streptomycin (100 μg/mL), 2-mercaptoethanol (2.5× 10−5 mol/L), and 10% fetal bovine serum (Invitrogen). Lewis lung carcinoma (LLC1) cells were obtained from ATCC in 2020 and maintained in DMEM containing 4mM L-glutamine, glucose (4500 mg/L), 1mM sodium pyruvate, sodium bicarbonate (1500 mg/L), and 10% FBS. Cells were routinely screened for Mycoplasma (LookOut® Mycoplasma PCR Detection kit, Sigma-Aldrich). For in vivo experiments, minimally passaged stock cells were freshly thawed and split once prior to implantation. Recombinant human IL15 was kindly provided by the Cancer Therapy Evaluation Program (NCI). InVivoMAB anti-mouse CD8a (Clone 53-6.7, used for CD8+ T-cell depletion), rat IgG2a isotype control, anti-mouse IFNAR-1 (Clone MAR1-5A3), and mouse IgG1 isotype control were all purchased from BioXCell (Lebanon, NH).
Pilones K.A., Charpentier M., Garcia-Martinez E., Daviaud C., Kraynak J., Aryankalayil J., Formenti S.C, & Demaria S. (2020). Radiotherapy cooperates with IL15 to induce antitumor immune responses. Cancer immunology research, 8(8), 1054-1063.
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4

RBC Transfusion and Immune Response

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Peripheral blood of KEL RBC mice was collected in 12% Citrate Phosphate Dextrose Adenine (CPDA-1, Jorgensen Labs, Melville, NY) by retro-orbital bleeding and leuko-reduced with a Pall (East Hills, NY) syringe filter. WT, IFNAR1−/−, or chimeric mice were transfused i.v. with 75 μL leuko-reduced packed RBCs, the mouse equivalent of 1 unit of human RBCs. For secondary responses, mice were re-transfused 28 days later. For IFNAR1 blocking experiments, WT mice were i.p. injected with 600 μg anti-mouse IFNAR1 (clone: MAR1-5A3, BioXCell, West Lebanon, NH), isotype control mouse IgG1 (MOPC-21, BioXCell) or PBS 24 hours prior to and 3 and 7 days after transfusion.
Gibb D.R., Liu J., Santhanakrishnan M., Natarajan P., Madrid D.J., Patel S., Eisenbarth S.C., Tormey C.A., Stowell S.R., Iwasaki A, & Hendrickson J.E. (2017). B cells require type 1 interferon to produce alloantibodies to transfused KEL-expressing RBCs in mice. Transfusion, 57(11), 2595-2608.
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