WB result of Phospho-IRF-3 (Ser386) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody incubation conditions: overnight at 4°C
Primary antibody: Phospho-IRF-3 (Ser386) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untransfected HT-29 whole cell lysate 20 µg
Lane 2: HT-29 transfected with 2.5 μg/ml Poly (I:C) for 6 hours whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 47 kDa
Observed MW: 55 kDa
Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Phospho-IRF3 (Ser386) |
| Synonyms | Interferon regulatory factor 3; IRF3 |
| Location | Cytoplasm, Nucleus, Mitochondrion |
| Accession | Q14653 |
| Clone Number | S-3498 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC |
| Reactivity | Hu |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu |
| ICC | 1:100 | Hu |
Background
Phospho-IRF3 (Ser386) refers to the activated form of interferon regulatory factor 3 (IRF3) phosphorylated at serine residue 386. IRF3 is a critical transcription factor in the innate immune response. Upon viral or bacterial infection, pattern recognition receptors (such as RIG-I, cGAS, or TLR3/4) in host cells recognize pathogen-associated molecular patterns, thereby activating the downstream kinases TBK1 and IKKε. These kinases directly phosphorylate multiple C-terminal serine/threonine residues of IRF3, among which phosphorylation of Ser386 is essential for IRF3 conformational change, dimerization, nuclear translocation, and transcriptional activity. Once phosphorylated at Ser386, IRF3 forms homodimers and translocates into the nucleus, where it binds to interferon-stimulated response elements (ISRE) and initiates the expression of type I interferons (such as IFN-β) and antiviral genes. Therefore, detecting the phosphorylation status of Ser386 is a common marker for monitoring IRF3 activation and assessing the level of innate immune responses, and is widely used in fields such as virology, autoimmune diseases, and vaccine development.
Picture
Picture
Western Blot
Immunocytochemistry
ICC analysis of HT-29 cells treated with poly(I:C) (2.5 µg/mL, 6 hr) (top panel) and untreated HT-29 cells (below panel). Anti- Phospho-IRF3 (Ser386) antibody was used at 1/100 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
