Knockout Tested Rabbit Recombinant Monoclonal IRF3 phospho S396 antibody. Carrier free. Suitable for Dot, WB and reacts with Synthetic peptide, Human samples.
Images
![Dot Blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (AB320083), expandable thumbnail](https://content.abcam.com/products/images/irf3-phospho-s396-antibody-epr28686-189-bsa-and-azide-free-ab320083--dot-blot-img435251.jpg/_jcr_content/renditions/webp-475.webp "Dot Blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (AB320083)")
![Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (AB320083), expandable thumbnail](https://content.abcam.com/products/images/irf3-phospho-s396-antibody-epr28686-189-bsa-and-azide-free-ab320083--western-blot-img435170.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (AB320083)")
![Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (AB320083), expandable thumbnail](https://content.abcam.com/products/images/irf3-phospho-s396-antibody-epr28686-189-bsa-and-azide-free-ab320083--western-blot-img435171.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (AB320083)")
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: 100% PBS- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| Dot | WB | ICC/IF | |
|---|---|---|---|
| Human | Expected | Tested | Not recommended |
| Mouse | Not recommended | Not recommended | Not recommended |
| Synthetic peptide | Tested | Not recommended | Not recommended |
Dot
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse, Synthetic peptide | Dilution info - | Notes - |
ICC/IF
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Mouse, Synthetic peptide | Dilution info - | Notes - |
Target data
Function
Key transcriptional regulator of type I interferon (IFN)-dependent immune responses which plays a critical role in the innate immune response against DNA and RNA viruses (PubMed:22394562, PubMed:24049179, PubMed:25636800, PubMed:27302953, PubMed:31340999, PubMed:36603579, PubMed:8524823). Regulates the transcription of type I IFN genes (IFN-alpha and IFN-beta) and IFN-stimulated genes (ISG) by binding to an interferon-stimulated response element (ISRE) in their promoters (PubMed:11846977, PubMed:16846591, PubMed:16979567, PubMed:20049431, PubMed:32972995, PubMed:36603579, PubMed:8524823). Acts as a more potent activator of the IFN-beta (IFNB) gene than the IFN-alpha (IFNA) gene and plays a critical role in both the early and late phases of the IFNA/B gene induction (PubMed:16846591, PubMed:16979567, PubMed:20049431, PubMed:36603579). Found in an inactive form in the cytoplasm of uninfected cells and following viral infection, double-stranded RNA (dsRNA), or toll-like receptor (TLR) signaling, is phosphorylated by IKBKE and TBK1 kinases (PubMed:22394562, PubMed:25636800, PubMed:27302953, PubMed:36603579). This induces a conformational change, leading to its dimerization and nuclear localization and association with CREB binding protein (CREBBP) to form dsRNA-activated factor 1 (DRAF1), a complex which activates the transcription of the type I IFN and ISG genes (PubMed:16154084, PubMed:27302953, PubMed:33440148, PubMed:36603579). Can activate distinct gene expression programs in macrophages and can induce significant apoptosis in primary macrophages (PubMed:16846591). In response to Sendai virus infection, is recruited by TOMM70:HSP90AA1 to mitochondrion and forms an apoptosis complex TOMM70:HSP90AA1:IRF3:BAX inducing apoptosis (PubMed:25609812). Key transcription factor regulating the IFN response during SARS-CoV-2 infection (PubMed:33440148).
Alternative names
Interferon regulatory factor 3, IRF-3, IRF3
Recommended products
Knockout Tested Rabbit Recombinant Monoclonal IRF3 phospho S396 antibody. Carrier free. Suitable for Dot, WB and reacts with Synthetic peptide, Human samples.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: 100% PBS- Form
- Liquid
- Clonality
- Monoclonal
- Immunogens
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
- Yes
- Clone number
- EPR28686-189
- Purification technique
- Affinity purification Protein A
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
Notes
ab320083 is the carrier-free version of Anti-IRF3 (phospho S396) antibody [EPR28686-189] ab320082.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free batch production
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
IRF3 also known as Interferon Regulatory Factor 3 acts as an important transcription factor in the immune response. It has a molecular weight of approximately 47 kDa. The IRF3 protein is mainly expressed in the cytoplasm and nucleus of various cell types including immune cells such as macrophages and dendritic cells. The protein becomes activated through phosphorylation a process frequently identified in its phosphorylated form phospo-IRF3 or p-IRF3 which facilitates its role in immune function.
- Biological function summary
IRF3 participates in the regulation of type I interferon (IFN) response a fundamental antiviral defense mechanism. IRF3 when phosphorylated forms a complex with CBP/p300 which then translocates to the nucleus to drive the expression of IFN-stimulated genes. This action strengthens the innate immune response and boosts the body's ability to counteract viral infections. Its activity and regulation are significant for maintaining a balanced immune response without excessive inflammation.
- Pathways
IRF3 is involved in the Toll-like receptor (TLR) and RIG-I-like receptor (RLR) signaling pathways both essential in pathogen recognition and response. Within these pathways IRF3 interacts with proteins such as MAVS and TBK1 to propagate immune signaling. The activation of IRF3 in these pathways results in the production of type I interferons and other cytokines orchestrating an effective antiviral response. These interactions highlight the protein's central role in mediating immune signaling cascades.
- Associated diseases and disorders
IRF3's malfunction or deregulation can contribute to autoimmune diseases and antiviral deficiencies. Conditions such as systemic lupus erythematosus (SLE) and chronic hepatitis B infection are linked to IRF3 activity. During autoimmune responses or viral persistence the aberrant activation of IRF3 can lead to inappropriate immune responses. The connection with proteins like STAT1 in these conditions highlights the complex network IRF3 engages in facilitating its impact on disease progression and immune dysregulation.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
![Dot Blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (ab320083), expandable thumbnail](https://content.abcam.com/products/images/irf3-phospho-s396-antibody-epr28686-189-bsa-and-azide-free-ab320083--dot-blot-img435251.jpg "Dot Blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (ab320083)")
Dot Blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (ab320083)
This data was developed using Anti-IRF3 (phospho S396) antibody [EPR28686-189] ab320082, the same antibody clone in a different buffer formulation.
Dot blot analysis of IRF3 (phospho S396) using Anti-IRF3 (phospho S396) antibody [EPR28686-189] ab320082 at 1:1000 (0.509 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Dot Blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] (Anti-IRF3 (phospho S396) antibody [EPR28686-189] ab320082) at 1/1000 dilution
Lane 1: Human IRF3 (phospho S396) peptide 1
Lane 2: Human IRF3 (phospho S396) peptide 2
Lane 3: Human IRF3 non-phospho peptide
Lane 4: Human IRF3 (phospho S398) peptide
SecondaryAll lanes: Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Exposure time: 180s
![Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (ab320083), expandable thumbnail](https://content.abcam.com/products/images/irf3-phospho-s396-antibody-epr28686-189-bsa-and-azide-free-ab320083--western-blot-img435170.jpg "Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (ab320083)")
Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (ab320083)
This data was developed using Anti-IRF3 (phospho S396) antibody [EPR28686-189] ab320082, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-IRF3 (phospho S396) antibody [EPR28686-189] ab320082 was shown to bind specifically to IRF3. Target of interest was observed at 60 kDa in wild-type Hela cell lysates (lane 2) with no signal observed at this size in IRF3 knockout cell line (lane 4) (lane 4, knockout cell line Human IRF3 knockout HeLa cell line ab255345 / knockout cell lysate Human IRF3 knockout HeLa cell lysate ab263784).
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-IRF3 antibody [EPR2418Y] (Anti-IRF3 antibody [EPR2418Y] ab68481) staining at 1/1000 dilution.
All lanes: Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] (Anti-IRF3 (phospho S396) antibody [EPR28686-189] ab320082) at 1/1000 dilution
Lane 1: Untreated wild-typeHeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 60 µg
Lane 2: Wild-type HeLa treated first with 80nM TPA for 24h, then change fresh medium, transfect 10ug/ml poly(dA:dT) for 2h whole cell lysate (untreated membrane) at 60 µg
Lane 3: Untreated IRF3 knockout HeLa whole cell lysate (untreated membrane) at 60 µg
Lane 4: IRF3 knockout HeLa treated first with 80nM TPA for 24h, then change fresh medium, transfect 10ug/ml poly(dA:dT) for 2h whole cell lysate (untreated membrane) at 60 µg
Lane 5: Untreated wild-typeHeLa whole cell lysate (alkaline phosphatase treated membrane) at 60 µg
Lane 6: Wild-type HeLa treated first with 80nM TPA for 24h, then change fresh medium, transfect 10ug/ml poly(dA:dT) for 2h whole cell lysate (alkaline phosphatase treated membrane) at 60 µg
Lane 7: Untreated IRF3 knockout HeLa whole cell lysate (alkaline phosphatase treated membrane) at 60 µg
Lane 8: IRF3 knockout HeLa treated first with 80nM TPA for 24h, then change fresh medium, transfect 10ug/ml poly(dA:dT) for 2h whole cell lysate (alkaline phosphatase treated membrane) at 60 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 60 kDa, 36 kDa
Exposure time: 92s
![Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (ab320083), expandable thumbnail](https://content.abcam.com/products/images/irf3-phospho-s396-antibody-epr28686-189-bsa-and-azide-free-ab320083--western-blot-img435171.jpg "Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (ab320083)")
Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] - BSA and Azide free (ab320083)
This data was developed using Anti-IRF3 (phospho S396) antibody [EPR28686-189] ab320082, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The identity of the bands between 25 kDa and 35 kDa are unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-IRF3 antibody [EPR2418Y] (Anti-IRF3 antibody [EPR2418Y] ab68481) staining at 1/1000 dilution.
All lanes: Western blot - Anti-IRF3 (phospho S396) antibody [EPR28686-189] (Anti-IRF3 (phospho S396) antibody [EPR28686-189] ab320082) at 1/1000 dilution
Lane 1: Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 60 µg
Lane 2: THP-1 treated first with 80nM TPA for 24h, then change fresh medium, transfect 10ug/ml poly(dA:dT) for 2h whole cell lysate at 60 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 60 kDa, 36 kDa
Exposure time: 48s
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