NR9456 cells cultured in eight-well chamber slides (Millipore) were fixed with 4% paraformaldehyde–PBS and permeabilized with 0.5% Triton X-100–PBS. Blocking and staining were performed with Duolink in situ PLA probes and detection reagents (Sigma). Fluorescence was analyzed with a BZ-X710-All-in-One fluorescence microscope (Keyence). Pictures were taken by using the Z-stacking function of the microscope and combined with a BZ-X analyzer. PLA dots were counted by ImageJ, and the values were normalized to the number of cells in the pictures. Thirty-three to 190 cells from 4 to 11 different pictures were analyzed for each condition. The primary antibodies used were anti-myc tag (Active Motif, Inc., 4E12), anti-AIM2 (Cell Signaling Technology, Inc., catalog number 13095), and anti-STING (Thermo Fischer Scientific PA5-23381 or Santa Cruz sc-241049) antibodies.
Anti sting
Manufactured by Thermo Fisher Scientific
Anti-STING is a laboratory reagent designed to inhibit the function of the STING (Stimulator of Interferon Genes) protein. STING is a key component of the innate immune response pathway, playing a role in the detection of cytosolic DNA and the subsequent activation of type I interferon production. The Anti-STING reagent can be used in research applications to study the STING signaling pathway and its involvement in various biological processes.
Automatically generated - may contain errors
2 protocols using anti sting
1
Proximity Ligation Assay for Protein Interactions
2
Investigating Type I IFN Signaling in Ischemia-Reperfusion Injury
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C57BL/6 wild‐type (WT) mice and interferon alpha receptor‐1 knockout (IFNAR1‐/‐) mice (male, aged 9–12 weeks, purchased from the Jackson Laboratory, Bar Harbor, ME) were used in the study. PDCA‐1 antibody (CD317 antibody, Miltenyl Biotec, Gaithersburg, MD) was used to deplete pDCs with rat IgG2b was used as isotype control. IFNAR1 monoclonal antibody (MAR1‐5A3, ThermoFisher Scientific, Grand Island, NY) was given as an 2 µg/g IV bolus 5 minutes before reperfusion to inhibit IFNAR1 activity. The signaling pathway leading to IFN‐I production was blocked using cyclic GMP‐AMP synthase (cGAS) inhibitor (RU.521 0.2 µg/g, InvivoGen), anti‐STING (stimulator of interferon genes) antibody (1 µg/g, ThermoFisher), anti‐IRF3 (interferon regulatory factor 3) antibody (1 µg/g, ThermoFisher) respectively given as intravenous boluses 5 minutes before reperfusion.
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