Product: Phospho-TBK1 (Ser172) Antibody
Catalog: AF8190
Description: Rabbit polyclonal antibody to Phospho-TBK1 (Ser172)
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 84kDa; 84kD(Calculated).
Uniprot: Q9UHD2
RRID: AB_2840252

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Product Info

Source:
Rabbit
Application:
WB 1:1000-3000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-TBK1 (Ser172) Antibody detects endogenous levels of TBK1 only when phosphorylated at Ser172.
RRID:
AB_2840252
Cite Format: Affinity Biosciences Cat# AF8190, RRID:AB_2840252.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

EC 2.7.11.1; FLJ11330; FTDALS4; NAK; NF kappa B activating kinase; NF kB activating kinase; NF-kappa-B-activating kinase; Serine/threonine protein kinase TBK 1; Serine/threonine protein kinase TBK1; Serine/threonine-protein kinase TBK1; T2K; TANK binding kinase 1; TANK-binding kinase 1; TBK 1; Tbk1; TBK1_HUMAN;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q9UHD2 TBK1_HUMAN:

Ubiquitous with higher expression in testis. Expressed in the ganglion cells, nerve fiber layer and microvasculature of the retina.

Sequence:
MQSTSNHLWLLSDILGQGATANVFRGRHKKTGDLFAIKVFNNISFLRPVDVQMREFEVLKKLNHKNIVKLFAIEEETTTRHKVLIMEFCPCGSLYTVLEEPSNAYGLPESEFLIVLRDVVGGMNHLRENGIVHRDIKPGNIMRVIGEDGQSVYKLTDFGAARELEDDEQFVSLYGTEEYLHPDMYERAVLRKDHQKKYGATVDLWSIGVTFYHAATGSLPFRPFEGPRRNKEVMYKIITGKPSGAISGVQKAENGPIDWSGDMPVSCSLSRGLQVLLTPVLANILEADQEKCWGFDQFFAETSDILHRMVIHVFSLQQMTAHKIYIHSYNTATIFHELVYKQTKIISSNQELIYEGRRLVLEPGRLAQHFPKTTEENPIFVVSREPLNTIGLIYEKISLPKVHPRYDLDGDASMAKAITGVVCYACRIASTLLLYQELMRKGIRWLIELIKDDYNETVHKKTEVVITLDFCIRNIEKTVKVYEKLMKINLEAAELGEISDIHTKLLRLSSSQGTIETSLQDIDSRLSPGGSLADAWAHQEGTHPKDRNVEKLQVLLNCMTEIYYQFKKDKAERRLAYNEEQIHKFDKQKLYYHATKAMTHFTDECVKKYEAFLNKSEEWIRKMLHLRKQLLSLTNQCFDIEEEVSKYQEYTNELQETLPQKMFTASSGIKHTMTPIYPSSNTLVEMTLGMKKLKEEMEGVVKELAENNHILERFGSLTMDGGLRNVDCL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Zebrafish
100
Chicken
100
Rabbit
100
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9UHD2 As Substrate

Site PTM Type Enzyme
K30 Ubiquitination
K60 Ubiquitination
K65 Ubiquitination
K69 Ubiquitination
K137 Ubiquitination
S151 Phosphorylation
Y153 Phosphorylation
K154 Ubiquitination
S172 Phosphorylation Q9UHD2 (TBK1)
Y174 Phosphorylation
Y179 Phosphorylation
K231 Ubiquitination
K236 Ubiquitination
K241 Ubiquitination
S247 Phosphorylation
K251 Ubiquitination
T278 Phosphorylation
K291 Ubiquitination
Y325 Phosphorylation
Y340 Phosphorylation
K344 Ubiquitination
Y354 Phosphorylation
K372 Ubiquitination
Y394 Phosphorylation
K396 Ubiquitination
K401 Ubiquitination
K416 Ubiquitination
Y435 Phosphorylation
K484 Ubiquitination
T503 Phosphorylation
K504 Ubiquitination
S509 Phosphorylation
S511 Phosphorylation
S527 Phosphorylation Q92630 (DYRK2)
K545 Ubiquitination
Y577 Phosphorylation
K584 Ubiquitination
K589 Ubiquitination
Y591 Phosphorylation
Y592 Phosphorylation
K596 Ubiquitination
K608 Ubiquitination
K615 Ubiquitination
K646 Ubiquitination
Y647 Phosphorylation
Y650 Phosphorylation
K661 Ubiquitination
T664 Phosphorylation
K670 Ubiquitination
T672 Phosphorylation
Y677 Phosphorylation
K702 Ubiquitination
S716 Phosphorylation

PTMs - Q9UHD2 As Enzyme

Substrate Site Source
O14920 (IKBKB) S177 Uniprot
O14920 (IKBKB) S181 Uniprot
P03372 (ESR1) S305 Uniprot
P06213 (INSR) S1033 Uniprot
P25963 (NFKBIA) S36 Uniprot
P31749 (AKT1) T308 Uniprot
P31749 (AKT1) S473 Uniprot
P42226 (STAT6) S407 Uniprot
P42226 (STAT6) Y641 Uniprot
Q01094 (E2F1) S332 Uniprot
Q04206 (RELA) S536 Uniprot
Q13501 (SQSTM1) S403 Uniprot
Q13568 (IRF5) S158 Uniprot
Q13568 (IRF5) S293 Uniprot
Q14653 (IRF3) S173 Uniprot
Q14653 (IRF3) S175 Uniprot
Q14653 (IRF3) S385 Uniprot
Q14653 (IRF3) S386 Uniprot
Q14653 (IRF3) S396 Uniprot
Q14653 (IRF3) S398 Uniprot
Q14653 (IRF3) S402 Uniprot
Q14653 (IRF3) T404 Uniprot
Q14653 (IRF3) S405 Uniprot
Q5S007 (LRRK2) S910 Uniprot
Q5S007 (LRRK2) S935 Uniprot
Q86WV6 (TMEM173) S358 Uniprot
Q92985 (IRF7) S471 Uniprot
Q92985 (IRF7) S472 Uniprot
Q92985 (IRF7) S477 Uniprot
Q92985 (IRF7) S479 Uniprot
Q96CV9 (OPTN) S177 Uniprot
Q96CV9 (OPTN) S473 Uniprot
Q96CV9 (OPTN) S513 Uniprot
Q96FA3 (PELI1) S76 Uniprot
Q96FA3 (PELI1) T80 Uniprot
Q96FA3 (PELI1) T288 Uniprot
Q96FA3 (PELI1) S293 Uniprot
Q9BRV8 (SIKE1) S133 Uniprot
Q9BRV8 (SIKE1) S185 Uniprot
Q9BRV8 (SIKE1) S187 Uniprot
Q9BRV8 (SIKE1) S188 Uniprot
Q9BRV8 (SIKE1) S190 Uniprot
Q9BRV8 (SIKE1) S198 Uniprot
Q9UHD2 (TBK1) S172 Uniprot

Research Backgrounds

Function:

Serine/threonine kinase that plays an essential role in regulating inflammatory responses to foreign agents. Following activation of toll-like receptors by viral or bacterial components, associates with TRAF3 and TANK and phosphorylates interferon regulatory factors (IRFs) IRF3 and IRF7 as well as DDX3X. This activity allows subsequent homodimerization and nuclear translocation of the IRFs leading to transcriptional activation of pro-inflammatory and antiviral genes including IFNA and IFNB. In order to establish such an antiviral state, TBK1 form several different complexes whose composition depends on the type of cell and cellular stimuli. Plays a key role in IRF3 activation: acts by first phosphorylating innate adapter proteins MAVS, STING1 and TICAM1 on their pLxIS motif, leading to recruitment of IRF3, thereby licensing IRF3 for phosphorylation by TBK1. Phosphorylated IRF3 dissociates from the adapter proteins, dimerizes, and then enters the nucleus to induce expression of interferons. Thus, several scaffolding molecules including FADD, TRADD, MAVS, AZI2, TANK or TBKBP1/SINTBAD can be recruited to the TBK1-containing-complexes. Under particular conditions, functions as a NF-kappa-B effector by phosphorylating NF-kappa-B inhibitor alpha/NFKBIA, IKBKB or RELA to translocate NF-Kappa-B to the nucleus. Restricts bacterial proliferation by phosphorylating the autophagy receptor OPTN/Optineurin on 'Ser-177', thus enhancing LC3 binding affinity and antibacterial autophagy. Phosphorylates SMCR8 component of the C9orf72-SMCR8 complex, promoting autophagosome maturation. Phosphorylates and activates AKT1. Seems to play a role in energy balance regulation by sustaining a state of chronic, low-grade inflammation in obesity, wich leads to a negative impact on insulin sensitivity (By similarity). Attenuates retroviral budding by phosphorylating the endosomal sorting complex required for transport-I (ESCRT-I) subunit VPS37C. Phosphorylates Borna disease virus (BDV) P protein. Plays an essential role in the TLR3- and IFN-dependent control of herpes virus HSV-1 and HSV-2 infections in the central nervous system.

PTMs:

Autophosphorylation at Ser-172 activates the kinase, and is an essential step for virus-triggered signaling. Phosphorylated by IKBKB/IKKB at Ser-172. Phosphorylation requires homodimerization and ubiquitination at Lys-30 and Lys-401. Dephosphorylated at Ser-172 by PPM1B and this negatively regulates its role in mediating antiviral response.

'Lys-63'-linked polyubiquitination by MIB1 after RNA virus infection, or by NRDP1 after LPS stimulation at Lys-30 and Lys-401, participates in kinase activation. 'Lys-48'-linked polyubiquitination at Lys-670 by DTX4 leads to proteasomal degradation. 'Lys-48'-linked polyubiquitination by TRAIP also leads to proteasomal degradation. 'Lys-63'-linked polyubiquitination by RNF128 at Lys-30 and Lys-401 leads to the activation of antiviral responses.

Subcellular Location:

Cytoplasm.
Note: Upon mitogen stimulation or triggering of the immune system, TBK1 is recruited to the exocyst by EXOC2.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Ubiquitous with higher expression in testis. Expressed in the ganglion cells, nerve fiber layer and microvasculature of the retina.

Subunit Structure:

Homodimer. Interacts with DDX3X, TIRAP and TRAF2. Part of a ternary complex consisting of TANK, TRAF2 and TBK1. Interacts with AZI2, TANK and TBKBP1; these interactions are mutually exclusive and mediate TBK1 activation. Interacts with GSK3B; this interaction promotes TBK1 self-association and autophosphorylation. Interacts with SIKE1; SIKE1 is associated with TBK1 under physiological condition and dissociated from TBK1 upon viral infection or TLR3 stimulation. Interacts with IRF3, leading to IRF3 phosphorylation. Interacts with DDX58/RIG-I. Interacts with CYLD. Interacts with OPTN and TRAF3. Interacts with SRC. Interacts with the exocyst complex subunit SEC5/EXOC2; this interaction is sufficient to trigger TBK1 activity. Interacts with STING1, leading to STING1 phosphorylation. Interacts with IFIT3 (via N-terminus). Interacts with MAVS; interaction only takes place in the presence of IFIT3 and leads to MAVS phosphorylation. Interacts (via protein kinase domain) with TTLL12 (via TTL domain); the interaction prevents MAVS binding to TBK1. Interacts with TICAM1; this interaction is enhanced in the presence of WDFY1 and leads to TICAM1 phosphorylation. Interacts with TRIM26. Interacts with TRIM23. Interacts with TTC4 and IKBKE. Interacts with HNRNPA2B1.

(Microbial infection) Interacts with Borna disease virus (BDV) P protein leading to its phosphorylation.

(Microbial infection) Interacts with Ebola virus protein VP35.

(Microbial infection) Interacts with HCV NS3; this interaction leads to inhibition of cellular antiviral response by blocking necessary interactions between the TBK1 and its substrates IRF3 and IRF7.

(Microbial infection) Interacts with herpes simplex virus 1 protein ICP34.5.

Family&Domains:

Comprises A N-terminal kinase domain, a ubiquitin-like domain and a C-terminal coiled-coil region mediating homodimerization.

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.

Research Fields

· Environmental Information Processing > Signal transduction > Ras signaling pathway.   (View pathway)

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Measles.

· Human Diseases > Infectious diseases: Viral > Influenza A.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Organismal Systems > Immune system > Toll-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > NOD-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > RIG-I-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Cytosolic DNA-sensing pathway.   (View pathway)

· Organismal Systems > Immune system > IL-17 signaling pathway.   (View pathway)

References

1). A Hybrid Nanoadjuvant Simultaneously Depresses PD-L1/TGF-β1 and Activates cGAS-STING Pathway to Overcome Radio-Immunotherapy Resistance. Advanced materials (Deerfield Beach, Fla.), 2024 (PubMed: 38229577) [IF=29.4]

2). CDK4/6 Alters TBK1 Phosphorylation to Inhibit the STING Signaling Pathway in Prostate Cancer. Cancer research, 2024 (PubMed: 38861362) [IF=11.2]

3). Commensal cow Roseburia reduces gut-dysbiosis-induced mastitis through inhibiting bacterial translocation by producing butyrate in mice. Cell Reports, 2022 (PubMed: 36417859) [IF=8.8]

4). Mechanism of selenomethionine inhibiting of PDCoV replication in LLC-PK1 cells based on STAT3/miR-125b5p-1/HK2 signaling. Frontiers in Immunology, 2022 (PubMed: 36059492) [IF=7.3]

5). Swine acute diarrhoea syndrome coronavirus (SADS-CoV) Nsp5 antagonizes type I interferon signaling by cleaving DCP1A. Frontiers in Immunology, 2023 (PubMed: 37283741) [IF=7.3]

6). TBK1 pharmacological inhibition mitigates osteoarthritis through attenuating inflammation and cellular senescence in chondrocytes. Journal of orthopaedic translation, 2024 (PubMed: 39040492) [IF=6.6]

7). TIPE2 knockout reduces myocardial cell damage by inhibiting IFN-γ-mediated ferroptosis. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE, 2023 (PubMed: 36216021) [IF=6.2]

8). The PICK1/TLR4 complex on microglia is involved in the regulation of LPS-induced sepsis-associated encephalopathy. International Immunopharmacology, 2021 (PubMed: 34500285) [IF=5.6]

Application: WB    Species: Mouse    Sample:

Fig. 5. Deficiency of PICK1 can lead to excessive activation of microglia and TLR4 signaling pathway in the hippocampus of mice with septic encephalopathy induced by LPS. (A) Representative images of Western blots of LPS-treated mice Hippocampus. (B–H) Quantitative analyses of PICK1, IBa1, TLR4, TRAF3, phosphoTBK1, phospho-IKKε, and phospho-IRF3.

9). Inhibition of TANK-binding kinase1 attenuates the astrocyte-mediated neuroinflammatory response through YAP signaling after spinal cord injury. CNS Neuroscience & Therapeutics, 2023 (PubMed: 37032635) [IF=5.5]

10). Wip1 Aggravates the Cerulein-Induced Cell Autophagy and Inflammatory Injury by Targeting STING/TBK1/IRF3 in Acute Pancreatitis. INFLAMMATION, 2021 (PubMed: 33417178) [IF=5.1]

Application: WB    Species: rat    Sample: pancreatic acinar cells

Fig. 3. Autophagy and STING pathway activation were involved in the development of AP. a Electron micrographs of pancreatic acinar cells in the control group showed that the lysosomes were very clear and there was no obvious phagocytosis. b The structure of pancreatic acinar cells was disordered in the AP group and more opaque substances were observed in the lysosomes, which were considered as autophagy lysosomes. c Western blot results showed that the level of P-STING/P-TBK1/P-IRF3 and LC3B II/I was significantly increased in the cerulein-treated rats. d Western blot results showed that the level of PSTING/P-TBK1/P-IRF3 and LC3B II/I was significantly increased in the cerulein-treated AR42J cells. *P < 0.05 compared with the control groups.

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