Anti-Phospho-IRAK1 (Thr209) Polyclonal Antibody 2 is a Rabbit antibody targeting Phospho-IRAK1 (Thr209). Anti-Phospho-IRAK1 (Thr209) Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human,Rat

1. Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (IRAK1 (Thr209)) Polyclonal Antibody, Unconjugated (TMAB-10870) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
2. Tissue/cell: Human lung cancer tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-Phospho-IRAK1 (Ser376) Polyclonal Antibody, Unconjugated (TMAB-10870) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
3. Sample: Raji Cell Lysate at 30 μg
Primary: Anti-phospho-IRAK1 (Thr209) (TMAB-10870) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 78 kD
Observed band size: 78 kD
4. Blank control (blue): RSC96 cells (fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice). Primary Antibody: Rabbit Anti-Phospho-IRAK1 (Thr209) antibody (TMAB-10870), Dilution: 0.2 μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions); Secondary Antibody: Goat anti-rabbit IgG-PE (white blue), Dilution: 1: 200 in 1 X PBS containing 0.5% BSA.
5. Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (Phospho-IRAK1 (Thr209)) polyclonal Antibody, Unconjugated (TMAB-10870) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100-500; IF: 1:100-500; FCM: 0.2μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human IRAK1 around the phosphorylation site of Thr209

Human

Irak1

Interleukin-1 receptor-associated kinase 1

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. Association with MYD88 leads to IRAK1 phosphorylation by IRAK4 and subsequent autophosphorylation and kinase activation. 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 the interferon regulatory factor 7 (IRF7) to induce its activation and translocation to the nucleus, resulting in transcriptional activation of type I IFN genes, which drive the cell in an antiviral state. When sumoylated, translocates to the nucleus and phosphorylates STAT3