Product Name: IRAK4-pT345+pS346
Product Number: AB-PK665
Size: 25 µg      Price:89.00
      $US
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: Protein kinase phosphosite-specific antibody

Antibody Code: PK665

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: T345+S346

Protein UniProt: Q9NWZ3

Protein SigNET: Q9NWZ3

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.

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.This antibody was also subject to negative purification over phosphotyrosine-agarose.

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: Human

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-pT345+pS346 blocking peptide

Related Product 2: IRAKSelectide - IRAK1 protein kinase substrate peptide

Scientific Background: IRAK4 is a protein-serine/threonine kinase of the TKL group and IRAK family .It plays a critical role in initiating innate immune response against foreign pathogens. It is involved in Toll-like receptor (TLR) and IL-1R signalling pathways, and is rapidly recruited by MYD88 to the receptor-signalling complex upon TLR activation 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.
Figure 1. Epitope mapping of IRAK4-pT345+pS346 antibody with similar phosphopeptides on dot blots.


Figure 2. Identification of phosphosites related to IRAK4-pT345+pS346.