Anti-IRAK1 Polyclonal Antibody for WB, IF/ICC, ELISA - P51617

Core Product Specifications and Parameters

Biological Background: IRAK1 Function and Localization

• IRAK1 (interleukin-1 receptor-associated kinase 1) is a serine/threonine kinase essential for initiating innate immune responses, acting as a key mediator in Toll-like receptor (TLR) and IL-1 receptor signaling pathways.
• Upon TLR/IL-1R activation, IRAK1 is rapidly recruited by MYD88, phosphorylated by IRAK4, and undergoes autophosphorylation, which triggers downstream signaling events.
• IRAK1 phosphorylates Pellino E3 ubiquitin ligases (PELI1, PELI2, PELI3), promoting polyubiquitination of IRAK1 itself, leading to recruitment of the IKK complex and subsequent NF-kB nuclear translocation.
• It also phosphorylates TIRAP to promote its ubiquitination and degradation, and directly phosphorylates IRF7 to drive type I interferon production, establishing an antiviral state.
• Subcellular localization includes cytoplasm, nucleus, and lipid droplets; sumoylation of IRAK1 facilitates nuclear translocation where it phosphorylates STAT3.
• Two main isoforms exist: isoform 1 is more strongly expressed than isoform 2, both ubiquitously present in human tissues.
• IRAK1 is involved in host-virus interactions and undergoes various post-translational modifications including phosphorylation, ubiquitination, and sumoylation.

Experimental Guidance and Technical Tips

• The immunogen corresponds to amino acids 424-633 of human IRAK1, a region conserved across human, mouse, and rat, supporting the observed cross-reactivity.
• For Western blot, consider using whole cell lysates from TLR/IL-1R-stimulated immune cells (e.g., THP-1, RAW264.7) to detect activation-dependent bands near 77 kDa; basal expression may vary.
• In IF/ICC experiments, optimize fixation and permeabilization conditions; endogenous IRAK1 may localize to both cytoplasm and nucleus depending on cell type and stimulation.
• For ELISA, pairing with a matched detection antibody (e.g., against a different epitope) may improve sensitivity; validate in the relevant sample matrix.
• As a polyclonal antibody, lot-to-lot variability is possible; include positive (e.g., stimulated cells) and negative controls in each assay.

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