The Toll-like receptor (TLR) family in mammals comprises a family of transmembrane proteins characterized by multiple copies of leucine rich repeats in the extracellular domain and IL-1 receptor motif in the cytoplasmic domain. Like its counterparts in Drosophila, TLRs signal through adaptor molecules. The TLR family is a phylogenetically conserved mediator of innate immunity that is essential for microbial recognition. Ten human homologs of TLRs (TLR1-10) have been described. TIRAP (TIR domain-containing adaptor protein) is an adaptor protein used by TLR4. Blocking TIRAP inhibits TLR4-mediated signaling events, including DC maturation and cytokine production.
Function:
Adapter involved in TLR2 and TLR4 signaling pathways in the innate immune response. Acts via IRAK2 and TRAF-6, leading to the activation of NF-kappa-B, MAPK1, MAPK3 and JNK, and resulting in cytokine secretion and the inflammatory response. Positively regulates the production of TNF-alpha and interleukin-6.
Subunit:
Homodimer. Also forms heterodimers with MyD88. Binds to TLR4 and IRAK2 via their respective TIR domains. Binds to BMX, PKR and TBK1. Does not interact with IRAK1, nor TLR9.
Subcellular Location:
Cytoplasm.
Tissue Specificity:
Highly expressed in liver, kidney, spleen, skeletal muscle and heart. Also detected in peripheral blood leukocytes, lung, placenta, small intestine, thymus, colon and brain.
Post-translational modifications:
Phosphorylated by IRAK1 and IRAK4. Also phosphorylated by BTK.
Polyubiquitinated. Polyubiquitination follows phosphorylation by BTK and leads to TIRAP degradation.
Similarity:
Contains 1 TIR domain.
SWISS:
P58753
Gene ID:
11929
Database links:
Entrez Gene: 11929 Human
Entrez Gene: 117149 Mouse
Entrez Gene: 136127 Rat
GenBank: AF378129 Human
GenBank: AF378130 Human
Omim: 606252 Human
SwissProt: P58753 Human
SwissProt: Q99JY1 Mouse
Unigene: 537126 Human
Unigene: 23987 Mouse
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Blank control:U937.
Primary Antibody (green line): Rabbit Anti-phospho-TIRAP(Tyr86) antibody (SL7562R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. 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.
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