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Rabbit anti irf3

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Rabbit anti-IRF3 is an antibody that recognizes the Interferon Regulatory Factor 3 (IRF3) protein. IRF3 is a key transcription factor involved in the induction of type I interferons in response to viral infection or other stimuli. This antibody can be used to detect and study the expression and activation of IRF3 in various experimental systems.

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Lab products found in correlation

Rabbit anti phospho irf3, by Cell Signaling Technology (4 mentions) Rabbit anti phospho tbk1, by Cell Signaling Technology (4 mentions) Rabbit anti tbk1, by Cell Signaling Technology (3 mentions) Rabbit anti tbk1, by Abcam (2 mentions) Mouse anti pcbp2, by Santa Cruz Biotechnology (2 mentions) Rabbit anti flag, by Merck Group (2 mentions) Rabbit anti sting, by Cell Signaling Technology (2 mentions) Mouse anti v5 antibody, by Immunoway (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Anti phospho irf3, by Cell Signaling Technology (1 mentions) Rig 1, by Merck Group (1 mentions) Ikk α, by Merck Group (1 mentions) Mouse monoclonal anti c myc, by Merck Group (1 mentions) Mouse anti β actin, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions) Goat anti mouse il 1β, by R&D Systems (1 mentions) Rabbit anti phospho irf3 ser396, by Cell Signaling Technology (1 mentions) Rabbit anti phospho stat1 tyr701, by Cell Signaling Technology (1 mentions) Rabbit anti mlkl phospho s345, by Abcam (1 mentions) Rabbit anti caspase 3, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Anti-IRF3 (65 citations) Rabbit anti-phospho-IRF3 (5 citations) Rabbit anti-phospho-IRF3 (Ser396) (4 citations) Rabbit anti-phospho-IRF3 (4D4G, (2 citations) Rabbit anti-phosphorylated IRF3 (Ser396) (2 citations) Rabbit anti-p-IRF3 (2 citations) Rabbit anti-IRF3 (D83B9, (2 citations)

10 protocols using rabbit anti irf3

1

Antibodies for Immune Signaling Pathway

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Rabbit anti-phospho-IRF3 (Cat# 4947, 1:1000), rabbit anti-IRF3 (Cat# 4302, 1:1000), rabbit anti-phospho-TBK1 (Cat# 5483, 1:1000) and rabbit anti-cGAS antibodies (Cat# 15102, Cat# 79978, Cat# 31659 (Mouse specific), 1:1000) were from Cell Signaling Technology. Rabbit anti-TBK1 (Cat# ab40676, 1:1000) was from Abcam. Rabbit anti-PCBP2 (Cat# 15070-1-AP, 1:1000) was from Proteintech. Mouse anti-PCBP2 (Cat# sc-101136, 1:1000) was from Santa Cruz Biotechnology. Mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Cat# KM9002, 1:4000), mouse anti-α-Tubulin (Cat# KM9007, 1:8000), and mouse anti-HA (Cat# KM8004, 1:1000) antibodies were from Sungene Biotechnology. Rabbit anti-Flag (Cat# F7425, 1:1000) antibody was from Sigma. Mouse anti-Flag (Cat# M185-3, 1:1000), rabbit anti-HA (Cat# M132-3, 1:1000), rabbit anti-Myc (Cat# 562, 1:1000), mouse anti-Myc (Cat# M192-3, 1:1000) antibodies were from MBL. Mouse anti-V5 antibody (Cat# YM3005, 1:1000) was from ImmunoWay Biotechnology Company. Rabbit anti-PCBP1 (Cat# A1044, 1:1000) antibody was from ABclonal Technology. Mouse/Rabbit anti-cGAS antibody was prepared in our laboratory, the cGAS antibody was generated by immunizing mice or rabbits with purified human cGAS full-length from E. coli.
Gu H., Yang J., Zhang J., Song Y., Zhang Y., Xu P., Zhu Y., Wang L., Zhang P., Li L., Chen D, & Sun Q. (2022). PCBP2 maintains antiviral signaling homeostasis by regulating cGAS enzymatic activity via antagonizing its condensation. Nature Communications, 13, 1564.
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2

Virus Infection Assay in PK-15 and HEK293T Cells

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Porcine kidney (PK-15) cells and cells of the human embryonic kidney cell line (HEK293T) were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% fetal bovine serum (Gibco) in a humidified atmosphere containing 5%CO2/95% air at 37°C. Virus infection experiments were carried out as previously described (35 (link)). Foot-and-mouth disease strain A/GDMM/CHA/2013 and Sendai virus (SeV) are maintained by State Key Laboratory.
Rabbit anti-IRF3 and anti-phospho-IRF3 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). RIG-I, TBK1, p-TBK1, IKK- α were acquired from Sigma-Aldrich. Alexa fluor-conjugated secondary antibodies for indirect immunofluorescence (IFA) were purchased from Life Technologies. Rabbit polyclonal anti-TPL2 and rabbit polyclonal anti-TPL2-T290 antibodies were obtained from Abcam. Mouse monoclonal anti-c-Myc and mouse monoclonal anti-c-Flag were purchased from Sigma Aldrich, and mouse anti-β-actin was bought from Thermo Fisher Scientific.
Hao J., Shen C., Wei N., Yan M., Zhang X., Xu G., Zhang D., Hou J., Cao W., Jin Y., Zhang K., Zheng H, & Liu X. (2021). Foot-and-Mouth Disease Virus Capsid Protein VP1 Antagonizes TPL2-Mediated Activation of the IRF3/IFN-β Signaling Pathway to Facilitate the Virus Replication. Frontiers in Immunology, 11, 580334.
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3

Immunoblotting of Cell Death Regulators

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The primary antibodies used for immunoblotting were mouse IgG1 anti-caspase-1 (p20) (Adipogen, #AG-20B-0042, 1/2000 dilution), mouse IgG2b anti-NLRP3 (Adipogen, #AG-20B-0014, 1/1000), goat anti-mouse IL-1β (R&D systems, #AF-401-NA, 1/1000), rabbit anti-GAPDH (Sigma-Aldrich, #G9545, 1/20,000), rabbit anti-IRF3 (Cell Signaling, #4302, 1/2000), rabbit anti-phospho-IRF3 (Ser396) (Cell Signaling, #4947, 1/2000), rabbit anti-MAVS (rodent specific) (Cell Signaling, # 4983, 1/1000), rabbit anti-phospho-STAT1 (Tyr701) (Cell Signaling, #9171, 1/1000), rabbit anti-STAT1 (D1K9Y) (Cell Signaling, #14994, 1/3000), rabbit anti-caspase-3 (Cell Signaling, #9662, 1/1000), mouse IgG1 anti-DDX33 (B-4) (Santa Cruz, #sc-390573, 1/1000), rabbit anti-MLKL (phospho S345) (Abcam, #ab196436, 1/1000), rabbit anti-MLKL (D6W1K) (Cell Signaling, #37705, 1/2000), rabbit anti-phospho-DRP1 (Ser616) (D9A1) (Cell Signaling, #4494, 1/1000), mouse IgG1 anti-DRP1 (BD Biosciences, #611113, 1/2000), rabbit anti-RIPK1 (D94C12) (Cell Signaling, #3493, 1/2000), and guinea pig anti-mouse gasdermin D (Adipogen, #AG-25B-0036, 1/1000).
da Costa L.S., Outlioua A., Anginot A., Akarid K, & Arnoult D. (2019). RNA viruses promote activation of the NLRP3 inflammasome through cytopathogenic effect-induced potassium efflux. Cell Death & Disease, 10(5), 346.
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4

Antibody Profiling for SARS-CoV-2 Analysis

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Antibodies used for immunoblot and immunofluorescence analysis include: rabbit anti-Flag (1:1000, Sigma), mouse anti-Flag M2 (1:1000, Sigma), rabbit anti-IRF3 (1:100 or 1:500, Cell Signaling Technology), mouse anti-IRF3 (1:1000, Cell Signaling Technology) rabbit anti-TBK1 (1:1000, Cell Signaling Technology), rabbit anti-phospho-TBK1 (1:1000, Cell Signaling Technology), rabbit anti-phospho-IRF3 (1:1000, Cell Signaling Technology), rabbit anti-IFIT1 (1:1000, Cell Signaling Technology), human anti-SARS-CoV-2 Spike (CR3022, 1:5000), rabbit anti-SARS-CoV-2 nucleocapsid (Genetex, 1:1000), rabbit anti-NF-κB (1:500, Cell Signaling Technology), rabbit anti-phospho-NF-κB (1:1000, Cell Signaling Technology), mouse anti-Strep (1:1000, Biolegend; 1:1000, Genscript), rabbit anti-Actin (1:1000, Cell Signaling Technology), Streptavidin-HRP (1:1000, Jackson ImmunoResearch), species-specific HRP-Conjugated antibodies (1:10,000, Jackson ImmunoResearch). Alexa Fluor conjugated secondary antibodies (1:500, Life Technologies), and nuclear counterstain 4’,6’-Diamidino-2-phenylindole dihydrochloride- (DAPI; 1:1000, ACROS Organics).
Vazquez C., Swanson S.E., Negatu S.G., Dittmar M., Miller J., Ramage H.R., Cherry S, & Jurado K.A. (2021). SARS-CoV-2 viral proteins NSP1 and NSP13 inhibit interferon activation through distinct mechanisms. PLoS ONE, 16(6), e0253089.
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5

Western Blot Analysis of STING Pathway

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At 24 hours post-transfection, growth media was aspirated, cells were carefully washed with PBS, and subsequently lysed with Cell Extraction Buffer (Life Tech) with added Pierce protease and phosphatase inhibitors. The lysates were incubated on ice for 15 min followed by centrifugation for 5 min at ≥8000 rcf and 4 °C. LDS Sample Buffer was added to cell lysates at a final concentration of 1× followed by incubation at 100 °C for 5 min. The lysates were then subjected to SDS-PAGE on NuPAGE™ 4%–12% Bis-Tris precast Protein Gels (Invitrogen). Proteins were then transferred from SDS-PAGE gel to polyvinylidene fluoride membrane and detected using the following primary antibodies: rabbit anti-STING (1:2000; Cell Signaling), rabbit anti-TBK1 (1:1000; Cell Signaling), and rabbit anti-IRF3 (1:1000; Cell Signaling), rabbit anti-cGAS (1:2000; Millipore), mouse anti-HA.11 (1:3000, Biolegend), monoclonal SV40 T-antigen antibody (1:1000 dilution; Pab416, Santa Cruz Biotech). Secondary antibodies used: anti-mouse IgGκ HRP (1:3000, 1:5000 for anti-HA.11; Santa Cruz Biotech) and mouse anti-rabbit HRP (1:3000; Santa Cruz Biotech). SuperSignal™ West Femto Maximum Sensitivity Substrate (Thermo Scientific) was used to visualize proteins following secondary antibody incubation.
Reus J.B., Trivino-Soto G.S., Wu L.I., Kokott K, & Lim E.S. (2020). SV40 Large T Antigen Is Not Responsible for the Loss of STING in 293T Cells but Can Inhibit cGAS-STING Interferon Induction. Viruses, 12(2), 137.
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6

Immunoblotting Antibody Validation for IFN-γ Signaling

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The antibodies were listed above for flow cytometry at in vitro coculture assay. For immunoblotting, the following antibodies were used: mouse anti–MHC-I (Santa Cruz Biotechnology, sc-55582), rabbit anti-pSTAT1 (Cell Signaling Technology, 7649), rabbit anti-STAT1 (CST, 14994), rabbit anti-IFNGR1 (Millipore, MABF753), mouse anti–β-actin (Sigma-Aldrich, A5441), rabbit anti–RIG-I (Cell Signaling Technology, 3743), rabbit anti–MDA-5 (Cell Signaling Technology, 5321), rabbit anti-MAVS (Cell Signaling Technology, 3993), rabbit anti-pIRF3 (Cell Signaling Technology, 29047), rabbit anti-IRF3 (Cell Signaling Technology, 4302), rabbit anti-p-p65 (Cell Signaling Technology, 3033), rabbit anti-p65 (Cell Signaling Technology, 8242), rabbit anti-pSTING (Cell Signaling Technology, 72971), and rabbit anti-STING (Cell Signaling Technology, 13647). rabbit anti-p65 (Cell Signaling Technology, 8242) was used for RIP and ChIP, and normal rabbit immunoglobulin G (IgG; Cell Signaling Technology, 2729) was served as a negative control.
Wang Y., Zhao Y., Guo W., Yadav G.S., Bhaskarla C., Wang Z., Wang X., Li S., Wang Y., Chen Y., Pattarayan D., Xie W., Li S., Lu B., Kammula U.S., Zhang M, & Yang D. (2022). Genome-wide gain-of-function screening characterized lncRNA regulators for tumor immune response. Science Advances, 8(49), eadd0005.
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7

Immunological Profiling of Autoantibodies

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Anti-nuclear antibodies including anti-dsDNA, -ssDNA, -nucleosomes or -histones were measured as previously described (22 (link)). Total IgM and IgG isotypes and cytokine/chemokine expression was quantified using Luminex-based assays (Affymetrix). Transcription factor analysis was accomplished using Nuclear Extraction and Procarta Transcription Factor Plex Kits according to manufacturer’s instructions (Affymetrix). Immunoblot analysis was performed as previously described (26 ) and the concentration of the primary antibodies were as follows: rabbit anti-IRF3 (1:1000, Cell Signaling), rabbit anti-IRF7 (1 mg/mL, Abcam) and mouse anti-GAPDH (1:500, US Biological) antibodies.
Cuda C.M., Misharin A.V., Gierut A.K., Saber R., Haines GK I.I.I., Hutcheson J., Hedrick S.M., Mohan C., Budinger G.S., Stehlik C, & Perlman H. (2014). Caspase-8 Acts as a Molecular Rheostat to Limit RIPK1- and MyD88-Mediated Dendritic Cell Activation. Journal of immunology (Baltimore, Md. : 1950), 192(12), 5548-5560.
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8

Western Blot Analysis of Immune Signaling

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The cells extracts were prepared by ultrasonication in RIPA buffer in the presence of a protease inhibitor cocktail (Sigma). Proteins in the fraction were separated into 7.5%, and 12% SDS-polyacrylamide gels. After transfer and blocking, the PVDF membranes were incubated at 4 °C overnight under gentle agitation with each primary antibody: goat anti-CatH (1:500), rabbit anti-toll-like receptor 3 (TLR3, 1:500, Santa Cruz), rabbit anti-phosphorylated interferon regulatory factor 3 (p-IRF3, 1:1000, Cell Signaling Technology), rabbit anti-IRF3 (1:1000, Cell Signaling Technology), and mouse anti-actin (1:5000; Abcam). After washing, the membranes were incubated with horseradish peroxidase (HRP)-labeled anti-goat (1:2000; GE Healthcare), anti-mouse (1:2000 R&D Systems) and anti-rabbit (1:2000; GE Healthcare) for 2 h at room temperature. Subsequently, the membrane-bound, HRP-labeled antibodies were detected using an enhanced chemiluminescence detection system (ECL kit; GE Healthcare) with an image analyzer (LAS-3000; Fuji Photo Film).
Ni J., Zhao J., Zhang X., Reinheckel T., Turk V, & Nakanishi H. (2021). Cathepsin H deficiency decreases hypoxia-ischemia-induced hippocampal atrophy in neonatal mice through attenuated TLR3/IFN-β signaling. Journal of Neuroinflammation, 18, 176.
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9

Western Blot Analysis of Innate Immunity Proteins

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Rabbit anti-TBK1 (Cat# 38066, 1:1000), rabbit anti-phospho-TBK1 (Cat# 5483, 1:1000), rabbit anti-IRF3 (Cat# 11904, 1:1000), rabbit anti-phospho-IRF3 (Cat# 37829, 1:1000), rabbit anti-LC3A/B (Cat# 12741, 1:1000) and rabbit anti-GAPDH (Cat# 5174, 1:1000) were from Cell Signaling Technology. Rabbit anti-HSV-1 (Cat# ab9533, 1:100) was from Abcam. Rabbit anti-SP1 (Cat# 21962-1-AP, 1:1000) and mouse anti-alpha tubulin (Cat# 66031-1-Ig, 1:1000) were from Proteintech. Mouse anti-TREX1 (Cat# sc-133112, 1:1000) was from Santa Cruz Biotechnology.
Wang X., Wang Y., Cao A., Luo Q., Chen D., Zhao W., Xu J., Li Q., Bu X, & Quan J. (2023). Development of cyclopeptide inhibitors of cGAS targeting protein-DNA interaction and phase separation. Nature Communications, 14, 6132.
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10

Antibodies for Immunological Signaling Pathways

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Rabbit anti-phospho-IRF3, rabbit anti-IRF3, rabbit anti-phospho-TBK1 and rabbit anti-cGAS antibodies were from Cell Signaling Technology. Rabbit anti-TBK1 was from Abcam. Rabbit anti-PCBP2 was from Abcam and Proteintech. Mouse anti-PCBP2 was from Santa Cruz Biotechnology. Mouse anti-glyceraldehyde-3phosphate dehydrogenase (GAPDH), mouse anti-α-Tublin and mouse anti-HA antibodies were from Sungene Biotechnology. Rabbit anti-Flag antibody was from Sigma. Mouse anti-Flag, rabbit anti-HA, rabbit anti-Myc, mouse anti-Myc antibodies were from MBL. Mouse/rabbit anti-cGAS antibody was prepared in our laboratory, the cGAS antibody was generated by immunizing mice or rabbits with puri ed human cGAS full-length from E. coli.
Gu H., Yang J., Zhang J., Zhang P., Li L., Chen D., & Sun Q. (2021). PCBP2 maintains antiviral signaling homeostasis by regulating cGAS enzymatic activity via antagonizing its condensation.
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