Interferon response factor 3 (IRF3) is a 50 kDa, 427 amino acid, member of the IRF family of proteins. Human IRF3 is a transcription factor that binds the interferon-sensitive response element (ISRE) and is activated by Toll-like receptors, TLR3 and TLR4. Viral entry or engagement of TLR3 or TLR4 stimulates IRF3 phosphorylation, nuclear translocation and stimulation of IFN production. There are potential alternate splice forms affecting IRF3 structure, one including a deletion of aa 201‑327, a second with the same deletion and an alternate Met(147) start site, and a third with a 125 aa substitution of the C-terminal 100 aa. Over the region used as immunogen, aa 206‑427, human IRF3 is 76% and 83% identical to the corresponding mouse and porcine proteins, respectively.
Key Product Details
Validated by
Species Reactivity
Applications
Label
Antibody Source
Product Specifications
Immunogen
Val206-Ser427
Accession # Q14653
Specificity
Clonality
Host
Isotype
Scientific Data Images for Human IRF3 Antibody
Detection of IRF3 in Daudi Human Cell Line by Flow Cytometry.
Daudi human Burkitt's lymphoma cell line was stained with Goat Anti-Human IRF3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4019, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
Detection of Human IRF3 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, U937 human histiocytic lymphoma cell line, Raji human Burkitt's lymphoma cell line, and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human IRF3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4019) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for IRF3 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
IRF3 in HeLa Human Cell Line.
IRF3 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human IRF3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4019) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human IRF3 by Simple WesternTM.
Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma parental cell line and IRF3 knockout HeLa cell line (KO), loaded at 0.2 mg/mL. A specific band was detected for IRF3 at approximately 57 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. Goat Anti-Human IRF3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4019) was used at 10 µg/mL followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Western Blot Shows Human IRF3 Specificity by Using Knockout Cell Line.
Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and IRF3 knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human IRF3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4019) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for IRF3 at approximately 50 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Applications for Human IRF3 Antibody
CyTOF-ready
Immunocytochemistry
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line
Intracellular Staining by Flow Cytometry
Sample: Daudi human Burkitt's lymphoma cell line fixed with paraformaldehyde and permeabilized with saponin
Knockout Validated
Simple Western
Sample: HeLa human cervical epithelial carcinoma cell line
Western Blot
Sample: HeLa human cervical epithelial carcinoma cell line, U937 human histiocytic lymphoma cell line, Raji human Burkitt's lymphoma cell line, and Daudi human Burkitt's lymphoma cell line
Flow Cytometry Panel Builder
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Advanced Features
- Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
- Spillover Popups - Visualize the spectra of individual fluorochromes
- Antigen Density Selector - Match fluorochrome brightness with antigen density
Formulation, Preparation, and Storage
Purification
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
Shipping
Stability & Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: IRF3
Long Name
Alternate Names
Gene Symbol
UniProt
Additional IRF3 Products
Product Documents for Human IRF3 Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human IRF3 Antibody
For research use only
Related Research Areas
Citations for Human IRF3 Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- 7-Amino Actinomycin D (7-AAD) Cell Viability Flow Cytometry Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Detection & Visualization of Antibody Binding
- Extracellular Membrane Flow Cytometry Protocol
- Flow Cytometry Protocol for Cell Surface Markers
- Flow Cytometry Protocol for Staining Membrane Associated Proteins
- Flow Cytometry Staining Protocols
- Flow Cytometry Troubleshooting Guide
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Intracellular Flow Cytometry Protocol Using Alcohol (Methanol)
- Intracellular Flow Cytometry Protocol Using Detergents
- Intracellular Nuclear Staining Flow Cytometry Protocol Using Detergents
- Intracellular Staining Flow Cytometry Protocol Using Alcohol Permeabilization
- Intracellular Staining Flow Cytometry Protocol Using Detergents to Permeabilize Cells
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Propidium Iodide Cell Viability Flow Cytometry Protocol
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Characterization of Human Th22 Cells
- Protocol for the Characterization of Human Th9 Cells
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- Protocol: Annexin V and PI Staining by Flow Cytometry
- Protocol: Annexin V and PI Staining for Apoptosis by Flow Cytometry
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Fluorokine Flow Cytometry Kits
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars