Product Name:
IRAK4-pT345+pS346
Product Number:
ab-pk665
Target Full Name: Interleukin 1 receptor-associated kinase 4
Target Alias: Interleukin-1 receptor associated kinase 4; IPD1, REN64, NY-REN-64; IRAK-4
Product Type Specific: IRAK4 protein kinase phosphosite-specific antibody
Antibody Code: PK665
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: T345+S346
Protein UniProt: Q9NWZ3
Protein SigNET: IRAK4
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human IRAK4 sequence peptide Cat. No.: PE-04ANA99
Antibody Immunogen Sequence: TVM(pT)(pS)RIV(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues T342 to V349; in the protein kinase catalytic domain activation T loop region between subdomains VII and VIII. T345 and S346 phosphorylation stimulate phosphotransferase activity. Phosphorylation regulates molecular association, including inducing interaction with Akt1. These are minor in vivo phosphorylation sites in IRAK4 (≥2 and ≥3 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). IRAK4 is known to be phosphorylated in vitro at T345 by IRAK4, and at S346 by IRAK4.
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Potency: Weak immunoreactivity of a target-sized protein by Western blotting in A431 cells. Very strong immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus macaque | Dog | Rat | Mouse.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 53-60 kDa.
Antibody Specificity: High-very high
Antibody Cross Reactivity: No significant cross-reactive proteins detected in Jurkat and A431 cells, exceptfor a ~77 kDa strongly cross-reactive protein detected in A431 cells.
Related Product 1: IRAK4 (342-349) pT345+pS346 antibody blocking peptide (Cat. No.: PE-04ANA99)
Related Product 2: IRAKSelectide kinase substrate peptide (Cat. No.: PE-01BHQ90)
Related Product 3: IRAK1 T387 phosphorylation antibody (Cat. No.: AB-PK664)
Scientific Background: IRAK4 is a 51-52 kDa protein-serine/threonine kinase of the TKL group and IRAK family. It functions as a crucial mediator in innate immunity, initiating signalling from Toll-like receptors (TLRs) and IL-1 receptors to activate NF-kB and pro-inflammatory pathways in response against foreign pathogens. TLR and IL-1R activation rapidly recruits MYD88 to the receptor-signalling complex to form the Myddosome together with IRAK2. It phosphorylates initially IRAK1, thus stimulating the kinase activity and intensive autophosphorylation of IRAK1. It phosphorylates E3 ubiquitin ligases Pellino proteins (PELI1, PELI2 and PELI3) to promote pellino-mediated polyubiquitination of IRAK1. Then, the ubiquitin-binding domain of IKBKG/NEMO binds to polyubiquitinated IRAK1 bringing together the IRAK1-MAP3K7/TAK1-TRAF6 complex and the NEMO-IKKA-IKKB complex. IRAK4 phosphorylates TIRAP to promote its ubiquitination and subsequent degradation. It phosphorylates NCF1 and regulates NADPH oxidase activation after LPS stimulation. Mutations in IRAK4 result in a nonfunctional protein, leading to severe susceptibility to pyogenic bacterial infections. Its overexpression is linked to inflammatory diseases, autoimmune disorders (e.g., rheumatoid arthritis), and cancers. This description may include information annotated by UniProt and/or Google AI.