Anti-Phospho-IRAK4 (Thr345) Polyclonal Antibody is a Rabbit antibody targeting Phospho-IRAK4 (Thr345). Anti-Phospho-IRAK4 (Thr345) Polyclonal Antibody can be used in WB,FCM.

IgG

Mouse (predicted:Human,Rat,Dog,Pig,Cow,Rabbit,Sheep,GuineaPig)

1. Sample: Lung (Mouse) Lysate at 40 μg
Primary: Anti-phospho-IRAK4 (Thr345) (TMAB-10874) at 1/300 dilution
Secondary: HRP conjugated Goat-Anti-rabbit IgG at 1/5000 dilution
Predicted band size: 51 kD
Observed band size: 60 kD
2. Blank control (black line): NIH/3T3.
Primary Antibody (green line): Rabbit Anti-phospho-IRAK4 (Thr345) antibody
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

WB: 1:500-2000; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human IRAK4 around the phosphorylation site of Thr345

Human

IRAK4

Interleukin-1 receptor-associated kinase 4

Serine/threonine-protein kinase that plays a critical role in initiating innate immune response against foreign pathogens. Involved in Toll-like receptor (TLR) and IL-1R signaling pathways. Is rapidly recruited by MYD88 to the receptor-signaling complex upon TLR activation to form the Myddosome together with IRAK2. Phosphorylates initially IRAK1, thus stimulating the kinase activity and intensive autophosphorylation of IRAK1. 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. In turn, MAP3K7/TAK1 activates IKKs (CHUK/IKKA and IKBKB/IKKB) leading to NF-kappa-B nuclear translocation and activation. Alternatively, phosphorylates TIRAP to promote its ubiquitination and subsequent degradation. Phosphorylates NCF1 and regulates NADPH oxidase activation after LPS stimulation suggesting a similar mechanism during microbial infections.