Myddosome Complex Antibody Sampler Kit #39433
Product Information
Kit Usage Information
Protocols
- 4283: Western Blotting, Immunoprecipitation (Magnetic)
- 4363: Western Blotting, Immunoprecipitation (Agarose)
- 4367: Western Blotting
- 4504: Western Blotting, Immunoprecipitation (Agarose), Flow
- 5483: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence, Flow
- 7074: Western Blotting
- 11927: Western Blotting, Immunoprecipitation (Magnetic)
- 38066: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence, Flow
- 67591: Western Blotting, Immunoprecipitation (Magnetic)
Product Description
Specificity / Sensitivity
Each antibody in the Myddosome Complex Antibody Sampler Kit detects endogenous levels of its target human protein. Phospho-IRAK4 (Thr345/Ser346) (D6D7) Rabbit mAb recognizes endogenous levels of IRAK4 protein when phosphorylated at Thr345 and Ser346. This antibody shows slight reactivity with IRAK4 when singly phosphorylated at Ser346 and does not cross-react with IRAK4 singly phosphorylated at Thr345. Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb detects endogenous levels of TBK1 only when phosphorylated at Ser172 and may cross-react with phospho-IKKε.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with recombinant protein specific to a central region of mouse TRAF6 proteins, with synthetic phosphopeptides corresponding to residues surrounding Thr345/Ser346 and Ser172 of human IRAK4 and TBK1/NAK1 proteins, respectively, and with synthetic peptides corresponding to residues surrounding Cys233 of human MyD88 protein and residues near the carboxyl termini of mouse IRAK1 and human TBK1/NAK protein. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues at the carboxy terminus of mouse IRAK2 and residues surrounding Lys41 of human IRAK4. Antibodies were purified by protein A and peptide affinity chromatography.
Background
Toll-like receptors (TLRs) are a large family of so-called pattern recognition receptors (PRRs) that detect pathogen associated molecular patterns (PAMPs) and danger associated molecular patterns (DAMPs) (1,2). Upon activation, TLRs initiate two main signaling pathways through their C-terminal cytoplasmic Toll/Il-1 receptor (TIR) domain that couples with TIR domain-containing adaptors MyD88 and TRIF. The MyD88-dependent pathway is initiated by the formation of a large oligomeric protein complex termed myddosome. Myddosome is one of so-called supramolecular organizing centers (SMOCs), a signaling organelle that is common for PRRs in the innate immune system. Myddosome formation promotes IRAK4 activation, which in turn activates IRAK1 and later, IRAK2. TRAF6 is then recruited and activated through the binding sites within IRAKs. Activated TRAF6 is released to the cytosol and triggers the IKK complex to activate the NF-κB pathway to mediate the expression of pro-inflammatory cytokines and chemokines (3-7). Recently, it was also found that TBK1 is recruited to the myddosome complex and activated to induce aerobic glycolysis (8).
- Behzadi, P. et al. (2021) J Immunol Res 2021, 9914854.
- Aluri, J. et al. (2021) Cells 10, 1374. doi: 10.3390/cells10061374.
- De Nardo, D. (2015) Cytokine 74, 181-9.
- Latty, S.L. et al. (2018) Elife 7:e31377. doi: 10.7554/eLife.31377.
- De Nardo, D. et al. (2018) J Biol Chem 293, 15195-15207.
- Balka, K.R. and De Nardo, D. (2019) J Leukoc Biol 105, 339-351.
- Gold, J.J. et al. (1988) Biosystems 21, 403-15.
- Tan, Y. and Kagan, J.C. (2019) Cell 177, 384-398.e11.
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