Phospho-IRF3 (Ser386) Antibody | Affinity Biosciences

Product:	Phospho-IRF3 (Ser386) Antibody
Catalog:	AF3438
Description:	Rabbit polyclonal antibody to Phospho-IRF3 (Ser386)
Application:	WB IHC IF/ICC
Cited expt.:	WB, IF/ICC
Reactivity:	Human, Rat
Prediction:	Pig, Bovine, Sheep, Dog
Mol.Wt.:	57kDa; 47kD(Calculated).
Uniprot:	Q14653
RRID:	AB_2834880

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

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Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Rat

Prediction:

Pig(92%), Bovine(92%), Sheep(92%), Dog(85%)

Clonality:

Polyclonal

Specificity:

Phospho-IRF3 (Ser386) Antibody detects endogenous levels of IRF3 only when phosphorylated at Serine 386.

RRID:

AB_2834880
Cite Format: Affinity Biosciences Cat# AF3438, RRID:AB_2834880.

Conjugate:

Unconjugated.

Purification:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

IIAE7; Interferon regulatory factor 3; IRF 3; IRF-3; IRF3; IRF3_HUMAN; MGC94729;

Immunogens

Immunogen:

A synthesized peptide derived from human IRF3 around the phosphorylation site of Ser386.

Uniprot:

Q14653(IRF3_HUMAN) >>Visit HPA database.

Gene(ID):

IRF3(3661)

Expression:

Q14653 IRF3_HUMAN:

Expressed constitutively in a variety of tissues.

Description:

IRF3 encodes interferon regulatory factor 3, a member of the interferon regulatory transcription factor (IRF) family. IRF3 is found in an inactive cytoplasmic form that upon serine/threonine phosphorylation forms a complex with CREBBP.

Sequence:

Q14653 IRF3_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MGTPKPRILPWLVSQLDLGQLEGVAWVNKSRTRFRIPWKHGLRQDAQQEDFGIFQAWAEATGAYVPGRDKPDLPTWKRNFRSALNRKEGLRLAEDRSKDPHDPHKIYEFVNSGVGDFSQPDTSPDTNGGGSTSDTQEDILDELLGNMVLAPLPDPGPPSLAVAPEPCPQPLRSPSLDNPTPFPNLGPSENPLKRLLVPGEEWEFEVTAFYRGRQVFQQTISCPEGLRLVGSEVGDRTLPGWPVTLPDPGMSLTDRGVMSYVRHVLSCLGGGLALWRAGQWLWAQRLGHCHTYWAVSEELLPNSGHGPDGEVPKDKEGGVFDLGPFIVDLITFTEGSGRSPRYALWFCVGESWPQDQPWTKRLVMVKVVPTCLRALVEMARVGGASSLENTVDLHISNSHPLSLTSDQYKAYLQDLVEGMDFQGPGES

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Pig

Bovine

Sheep

Dog

Horse

Chicken

Xenopus

Zebrafish

Rabbit

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

Q14653_IRF3_HUMAN

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. 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. 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. 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. 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. Can activate distinct gene expression programs in macrophages and can induce significant apoptosis in primary macrophages.

PTMs:

Constitutively phosphorylated on many Ser/Thr residues. Activated following phosphorylation by TBK1 and IKBKE. Innate adapter protein MAVS, STING1 or TICAM1 are first activated by viral RNA, cytosolic DNA, and bacterial lipopolysaccharide (LPS), respectively, leading to activation of the kinases TBK1 and IKBKE. These kinases then phosphorylate the adapter proteins on the pLxIS motif, leading to recruitment of IRF3, thereby licensing IRF3 for phosphorylation by TBK1. Phosphorylated IRF3 dissociates from the adapter proteins, dimerizes, and then enters the nucleus to induce IFNs.

(Microbial infection) Phosphorylation and subsequent activation of IRF3 is inhibited by vaccinia virus protein E3.

Ubiquitinated; ubiquitination involves RBCK1 leading to proteasomal degradation. Polyubiquitinated; ubiquitination involves TRIM21 leading to proteasomal degradation.

ISGylated by HERC5 resulting in sustained IRF3 activation and in the inhibition of IRF3 ubiquitination by disrupting PIN1 binding. The phosphorylation state of IRF3 does not alter ISGylation.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Shuttles between cytoplasmic and nuclear compartments, with export being the prevailing effect (PubMed:10805757). When activated, IRF3 interaction with CREBBP prevents its export to the cytoplasm (PubMed:10805757).

Tissue Specificity:

Expressed constitutively in a variety of tissues.

Family&Domains:

Belongs to the IRF family.

Research Fields

· Human Diseases > Infectious diseases: Bacterial > Pertussis.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Measles.

· Human Diseases > Infectious diseases: Viral > Influenza A.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Organismal Systems > Immune system > Toll-like receptor signaling pathway. (View pathway)

· Organismal Systems > Immune system > NOD-like receptor signaling pathway. (View pathway)

· Organismal Systems > Immune system > RIG-I-like receptor signaling pathway. (View pathway)

· Organismal Systems > Immune system > Cytosolic DNA-sensing pathway. (View pathway)

References

1). RIG-I, a novel DAMPs sensor for myoglobin activates NF-κB/caspase-3 signaling in CS-AKI model. Military Medical Research, 2021 (PubMed: 34148549) [IF=16.7]

2). Pharmacological manipulation of TRPC5 by kaempferol attenuates metastasis of gastrointestinal cancer via inhibiting calcium involved in the formation of filopodia. International journal of biological sciences, 2024 (PubMed: 39309444) [IF=8.2]

Application: IF/ICC Species: human Sample: MKN-45 and DLD-1 cells

Figure 6. Ca2+-dependent MLC activation enhances filopodia formation via p-cortactin rearrangement. (A) Representative immunofluorescence (IF) images of MKN-45 and DLD-1 cells stained for mitochondria and F-actin (red: mitochondria, green: F-actin). Scale bar: 10 μm. (B) Representative MYO10 IF staining in MKN-45 and DLD-1 cells treated with CCCP (10 μM) (blue: nuclei, red: MYO10, green: F-actin). Scale bar: 20 μm. (C) Quantification of MYO10-positive filopodia in MKN-45 and DLD-1 cells treated with CCCP (10 μM). Data are expressed as means ± SD, **P < 0.01, ***P < 0.001 (vs. DMSO group). (D) Representative IF staining of MKN-45 and DLD-1 cells (blue: nuclei, red: dil, green: MYO10, violet: p-cortactin). Scale bar: 20 μm (left). Zoomed-in images and fluorescence co-localization of p-cortactin and MYO10 at filopodia tips in MKN-45 and DLD-1 cells (MKN-45: Pearson > 0.9; DLD-1: Pearson > 0.9). Scale bar: 2 μm (right). (E) Quantification of p-cortactin-positive filopodia tips per cell in MKN-45 and DLD-1 cells treated with CaCl2 (5 mM) or BAPTA-AM (10 μM). Data are expressed as means ± SD, *P < 0.05, **P < 0.01 (vs. DMSO group). (F) Quantification of p-cortactin-positive filopodia tips per cell in MKN-45 and DLD-1 cells treated with calyculin A (50 nM) or P18 (8 μM). Data are expressed as means ± SD, *P < 0.05, **P < 0.01, ***P < 0.001 (vs. DMSO group). (G) Representative MYO10 IF staining in MKN-45 and DLD-1 cells treated with PP2 (10 μM) (blue: nuclei, red: MYO10, green: F-actin). Scale bar: 20 μm. (H) Quantification of MYO10-positive filopodia in MKN-45 and DLD-1 cells treated with PP2 (10 μM). Data are expressed as means ± SD, *P < 0.05, **P < 0.01 (vs. DMSO group).

3). FAM210B activates STAT1/IRF9/IFIT3 axis by upregulating IFN-α/β expression to impede the progression of lung adenocarcinoma. Cell death & disease, 2025 (PubMed: 39900908) [IF=8.1]

4). HAO1-mediated oxalate metabolism promotes lung pre-metastatic niche formation by inducing neutrophil extracellular traps. Oncogene, 2022 (PubMed: 35739335) [IF=6.9]

Application: WB Species: Mouse Sample: alveolar epithelial cells

Fig. 6: TLR3 signal activation induces HAO1 expression in alveolar epithelial cells. A Effect of 67NR and 4T1 exosomes treatment on HAO1 expression in alveolar epithelial cells by Western blot analysis. B Effect of transfection of RNA isolated from 67NR and 4T1 exosomes on HAO1 expression in alveolar epithelial cells by Western blot analysis. C Effect of poly (I:C) treatment on HAO1 expression in alveolar epithelial cells by Western blot analysis. D Effect of poly (I:C) inhalation on HAO1 expression in mice lung by Western blot analysis. E Detection of oxalate production in alveolar epithelial cells treated with poly(I:C) or poly(I:C) + CCPST. Means ± s.e.m are provided (n = 3). F Detection of TLR3 expression in alveolar epithelial cells infected with sg-control or sg-Tlr3 lentivirus. G Detection of HAO1 expression in alveolar epithelial cells treated with 4T1 exosomes, 4T1 exosomes + CU CPT 4a or 4T1 exosomes + sg-Tlr3 lentivirus by Western blot analysis. H Detection of oxalate production in alveolar epithelial cells treated with 4T1 exosomes, 4T1 exosomes + CU CPT 4a or 4T1 exosomes + sg-Tlr3 lentivirus. I Effect of IRF3 over-expression on IRF3, P-IRF3 and HAO1 expression in alveolar epithelial cells by Western blot analysis. J Diagram of the HAO1 promoter showing the location of IRF3 binding sites. K Luciferase activity of HAO1-promoter construct after transfection of IRF3 plasmid in alveolar epithelial cells. L Detection of IRF3, P-IRF3 and HAO1 expression in alveolar epithelial cells treated with 4T1 exosomes or 4T1 exosomes + shIRF3 by Western blot analysis. M Detection of oxalate production in alveolar epithelial cells treated with 4T1 exosomes or 4T1 exosomes + shIRF3. Means ± s.e.m are provided (n = 3). N Schematic diagram of the role of HAO1-mediated oxalate metabolism at pre-metastatic stage and metastatic stage. ***P

5). Inhibition of Stimulator of Interferon Genes Protects Against Myocardial Ischemia-Reperfusion Injury in Diabetic Mice. Cardiovascular Innovations and Applications, 2023 [IF=0.9]

Application: WB Species: Mouse Sample: Hearts

Figure 4 STING Knockout Mitigates the Inflammatory Response in Diabetic Mice Subjected to I/R Injury. Mice were treated as described in Figure 2. (A) Representative western blotting results of p-TBK, p-IRF3, and IL-1β. (B) Statistical analysis of p-TBK/TBK in each group. (C) Statistical analysis of p-IRF3/IRF3 in each group. (D) Statistical analysis of IL-1β/GAPDH in each group. (E) Immunohistochemical results of TNF-α and IL-1β. Data are expressed as means ± SD, n=3.

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Phospho-IRF3 (Ser386) Antibody - #AF3438