Product: TLR4 Antibody
Catalog: AF7017
Source: Rabbit
Application: WB, IHC, IF/ICC, ELISA(peptide)
Reactivity: Human, Mouse, Rat
Mol.Wt.: 100~130kD; 96kD(Calculated).
Uniprot: O00206
RRID: AB_2835322

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Product Info

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.
TLR4 Antibody detects endogenous levels of total TLR4.
Cite Format: Affinity Biosciences Cat# AF7017, RRID:AB_2835322.
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
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.


ARMD10; CD284; CD284 antigen; Homolog of Drosophila toll; hToll; TLR 4; TLR4; TLR4_HUMAN; TOLL; Toll like receptor 4; Toll-like receptor 4;


O00206 TLR4_HUMAN:

Highly expressed in placenta, spleen and peripheral blood leukocytes (PubMed:9435236, PubMed:9237759). Detected in monocytes, macrophages, dendritic cells and several types of T-cells (PubMed:9237759, PubMed:27022195).

Toll like receptors (TLR) are highly conserved throughout evolution and have been implicated in the innate defense to many pathogens. In Drosophila toll is required for the anti fungal response, while the related 18 wheeler is involved in antibacterial defenses. In mammals, TLR identified as type I transmembrane signaling receptors with pattern recognition capabilities have been implicated in the innate host defense to pathogens. TLR4 has been identified next to MD2 and CD14 as a receptor that is central to the innate immune response to lipopolysaccharides (LPS) of Gram negative bacteria.



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.

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O00206 As Substrate

Site PTM Type Enzyme
N35 N-Glycosylation
N173 N-Glycosylation
T175 Phosphorylation
N205 N-Glycosylation
N282 N-Glycosylation
N309 N-Glycosylation
N497 N-Glycosylation
N526 N-Glycosylation
Y551 Phosphorylation
N575 N-Glycosylation
N624 N-Glycosylation
Y674 Phosphorylation
Y680 Phosphorylation
S730 Phosphorylation
S738 Phosphorylation
S790 Phosphorylation Q15139 (PRKD1)
S800 Phosphorylation

Research Backgrounds


Cooperates with LY96 and CD14 to mediate the innate immune response to bacterial lipopolysaccharide (LPS). Acts via MYD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Also involved in LPS-independent inflammatory responses triggered by free fatty acids, such as palmitate, and Ni(2+). Responses triggered by Ni(2+) require non-conserved histidines and are, therefore, species-specific. Both M.tuberculosis HSP70 (dnaK) and HSP65 (groEL-2) act via this protein to stimulate NF-kappa-B expression. In complex with TLR6, promotes sterile inflammation in monocytes/macrophages in response to oxidized low-density lipoprotein (oxLDL) or amyloid-beta 42. In this context, the initial signal is provided by oxLDL- or amyloid-beta 42-binding to CD36. This event induces the formation of a heterodimer of TLR4 and TLR6, which is rapidly internalized and triggers inflammatory response, leading to the NF-kappa-B-dependent production of CXCL1, CXCL2 and CCL9 cytokines, via MYD88 signaling pathway, and CCL5 cytokine, via TICAM1 signaling pathway, as well as IL1B secretion. Binds electronegative LDL (LDL(-)) and mediates the cytokine release induced by LDL(-). Stimulation of monocytes in vitro with M.tuberculosis PstS1 induces p38 MAPK and ERK1/2 activation primarily via TLR2, but also partially via this receptor.


N-glycosylated. Glycosylation of Asn-526 and Asn-575 seems to be necessary for the expression of TLR4 on the cell surface and the LPS-response. Likewise, mutants lacking two or more of the other N-glycosylation sites were deficient in interaction with LPS.

Phosphorylated on tyrosine residues by LYN after binding lipopolysaccharide.

Subcellular Location:

Cell membrane>Single-pass type I membrane protein. Early endosome. Cell projection>Ruffle.
Note: Upon complex formation with CD36 and TLR6, internalized through dynamin-dependent endocytosis (PubMed:20037584). Colocalizes with RFTN1 at cell membrane and then together with RFTN1 moves to endosomes, upon lipopolysaccharide stimulation.

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

Highly expressed in placenta, spleen and peripheral blood leukocytes. Detected in monocytes, macrophages, dendritic cells and several types of T-cells.

Subunit Structure:

Belongs to the lipopolysaccharide (LPS) receptor, a multi-protein complex containing at least CD14, LY96 and TLR4. Binding to bacterial LPS leads to homodimerization. Interacts with LY96 via the extracellular domain. Interacts with MYD88 and TIRAP via their respective TIR domains (By similarity). Interacts with TICAM2. Interacts with NOX4. Interacts with CNPY3 (By similarity). Interacts with HSP90B1. The interaction with both CNPY3 and HSP90B1 is required for proper folding in the endoplasmic reticulum. Interacts with MAP3K21; this interaction leads to negative regulation of TLR4 signaling. Interacts with CD36, following CD36 stimulation by oxLDL or amyloid-beta 42, and forms a heterodimer with TLR6. The trimeric complex is internalized and triggers inflammatory response. LYN kinase activity facilitates TLR4-TLR6 heterodimerization and signal initiation. Interacts with TICAM1 in response to LPS in a WDFY1-dependent manner. Interacts with WDFY1 in response to LPS (By similarity). Interacts with SMPDL3B (By similarity). Interacts with CEACAM1; upon lipopolysaccharide stimulation, forms a complex including TLR4 and the phosphorylated form of SYK and CEACAM1, which in turn, recruits PTPN6 that dephosphorylates SYK, reducing the production of reactive oxygen species (ROS) and lysosome disruption, which in turn, reduces the activity of the inflammasome (By similarity). Interacts with RFTN1; the interaction occurs in response to lipopolysaccharide stimulation. Interacts with SCIMP; the interaction occurs in response to lipopolysaccharide stimulation and is enhanced by phosphorylation of SCIMP by LYN (By similarity). This interaction facilitates the phosphorylation of TLR4 by LYN which elicits a selective cytokine response in macrophages (By similarity).

(Microbial infection) In case of infection, interacts with uropathogenic E.coli protein TcpC.


The TIR domain mediates interaction with NOX4.

The TIR domain mediates NAD(+) hydrolase (NADase) activity. Self-association of TIR domains is required for NADase activity.

Belongs to the Toll-like receptor family.

Research Fields

· Cellular Processes > Transport and catabolism > Phagosome.   (View pathway)

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Human Diseases > Infectious diseases: Bacterial > Pathogenic Escherichia coli infection.

· Human Diseases > Infectious diseases: Bacterial > Salmonella infection.

· Human Diseases > Infectious diseases: Bacterial > Pertussis.

· Human Diseases > Infectious diseases: Bacterial > Legionellosis.

· Human Diseases > Infectious diseases: Parasitic > Leishmaniasis.

· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).

· Human Diseases > Infectious diseases: Parasitic > Malaria.

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Parasitic > Amoebiasis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

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

· Human Diseases > Infectious diseases: Viral > Measles.

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

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Immune diseases > Inflammatory bowel disease (IBD).

· Human Diseases > Immune diseases > Rheumatoid arthritis.

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

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


1). Furuya DT et al. Anti-inflammatory effect of piperine ameliorates insulin resistance in monosodium glutamate–treated obese mice. Metabolism 2010 Mar;59(3):395-9. (PubMed: 19800637) [IF=21.567]

2). Wang J et al. Integrin 51, as a Receptor of Fibronectin, Binds the FbaA Protein of Group A Streptococcus To Initiate Autophagy during Infection. mBio 2020 Jun 9;11(3):e00771-20. (PubMed: 32518187) [IF=6.784]

Application: WB    Species: Mouse    Sample: Hep2 cells

FIG 6 FbaA initiates autophagy upon Fn-mediated binding to integrin 51. (A and B) Western blot analysis of TLR2 (A) and TLR4 (B) expression in Hep2 cells stimulated with FbaA for the indicated times. (C and D) Expression of LC3II in Hep2 cells with and without TLR2- or TLR4-specific siRNA following FbaA stimulation. (E) Schematic representation of the Fn subunit. Fn contains domain types I, II, and III. FnI is the region critical for binding to FnBP; FnIII9 and FnIII10 contain motifs that interact with integrins. (F) Hep2 cells were transfected with GST-FbaA, GST-FbaAFn, or GST (as a control), and the integrin 5 or 1 chain was measured after being pulled down with anti-GST Ab. (G) Expression of Fn, integrin 5, integrin 1, and LC3II in Hep2 cells stimulated with FbaA at the indicated concentrations, as analyzed by Western blotting. (H) Western blot analysis of LC3II expression in Hep2 cells stimulated with different concentrations of Fn and mock-FbaA or FbaA. (I to K) Expression of LC3II in Hep2 cells with or without specific siRNA against Fn, integrin 5, or integrin 1 following FbaA stimulation, as analyzed by Western blotting. (L and M) Fn, integrin 5, and integrin 1 knockdown cells were pretreated without (L) or with (M) rapamycin for 0.5 h and then infected with M1 GAS strain SF370 for 6 h. Numbers of CFU of M1 GAS strain SF370 were counted in the cells. (N) M1 GAS strain SF370-infected sh-Atg5-Hep2 cells with specific siRNA against Fn, integrin 5, or integrin 1. Numbers of CFU of M1 GAS strain SF370 were counted in the cells. *, P  0.05.

Application: WB    Species: human    Sample: Hep2 cells

FIG 6 |FbaA initiates autophagy upon Fn-mediated binding to integrin 51. (A and B) Western blot analysis of TLR2 (A) and TLR4 (B) expression in Hep2 cells stimulated with FbaA for the indicated times.

3). Shen Y et al. Inhibition of murine herpesvirus-68 replication by IFN-gamma in macrophages is counteracted by the induction of SOCS1 expression. PLoS Pathog 2018 Aug 3;14(8):e1007202 (PubMed: 30075008) [IF=6.218]

Application: WB    Species: mouse    Sample: macrophages

Fig 6. |TLR3 mediates the MHV-68-induced SOCS1 production. (A) BMMs were transfected with siRNAs against Tlr2, Tlr3, Tlr4, Tlr7, Tlr9 (si-TLR) or an irrelevant target (si-Control), respectively. Forty-eight hours post transfection, cells were infected with MHV-68 at MOI = 10. At 8 hpi, RNA and protein were then extracted. RT-qPCR was performed to determine SOCS1 mRNA levels and western blotting was performed to determine the protein expression levels of SOCS1 and the individual TLRs, respectively. The SOCS1 mRNA levels are expressed as values relative to the MHV-68 infected, si-Control transfected cells.

4). Zhou Y et al. Berberine-mediated up-regulation of surfactant protein D facilitates cartilage repair by modulating immune responses via the inhibition of TLR4/NF-ĸB signaling. Pharmacol Res 2020 Feb 11:104690 (PubMed: 32057894) [IF=5.893]

Application: WB    Species: rat    Sample: knee joints

Fig. 5. Chondrocyte apoptosis and immune molecular biomarkers were inhibited by BBR binding to SP-D in vivo in OA. OA model rats (n = 5/group) were injected using 25 μl of 200 μM BBR, followed by 25 μl rhSP-D (40 μg/mL) 1 day later. Sham-operated and OA-induction groups received an injection of 50 μl PBS into the right knee joint. For rats receiving rAAV-GFP or rAAV-SP-D shRNA, injections were performed for 10 consecutive days beginning 1 day after the initial BBR injection. At 10 weeks post-surgery, animals were sacrificed and the knee joints were collected for TUNEL and IHC analysis. (A) TUNEL staining (400 ×) was used to assess apoptosis. The p-p65, TLR4, F4/80, CD68, and CD34 expression was assessed via IHC (400 × with higher 800 × magnification inset). (B) Quantification of the indicated proteins in IHC images. Each column represents the mean ± SEM (n = 5). ***P < 0.001 versus the sham-operated group; ##P < 0.01 and ###P < 0.001 versus the OA-induction group; &P < 0.05 and &&P < 0.01 versus the OA + BBR group.

5). Li C et al. Oxyberberine, a novel gut microbiota-mediated metabolite of berberine, possesses superior anti-colitis effect: Impact on intestinal epithelial barrier, gut microbiota profile and TLR4-MyD88-NF-κB pathway. Pharmacol Res 2020 Feb;152:104603 (PubMed: 31863867) [IF=5.893]

Application: WB    Species: Mice    Sample: colonic tissues

Fig. 6. Effect of OBB on the activation of TLR4-MyD88-NF-κB signaling pathway in DSS-induced colonic tissues. (A) Representative Western blotting images of TLR4, MyD88, cytoplasmic p65, nuclear p65, p-IκBα and IκBα. Changes in the relative protein expression levels of TLR4 (B), MyD88 (C), nuclear p65 (D), cytoplasmic p65 (E), and p-IκBα/IκBα ratio (F) were measured. Data are shown as the mean ± SEM (n = 3). # P < 0.05, ## P < 0.01 vs. Control group, * P < 0.05, ** P < 0.01 vs. DSS group.

6). Zhang C et al. Extracellular vesicles derived from mesenchymal stem cells alleviate neuroinflammation and mechanical allodynia in interstitial cystitis rats by inhibiting NLRP3 inflammasome activation. J Neuroinflammation 2022 Apr 6;19(1):80. (PubMed: 35387668) [IF=5.793]

7). An K et al. Arsenic trioxide ameliorates experimental autoimmune encephalomyelitis in C57BL/6 mice by inducing CD4+ T cell apoptosis. J Neuroinflammation 2020 May 6;17(1):147. (PubMed: 32375831) [IF=5.793]

8). Guo LT et al. Baicalin ameliorates neuroinflammation-induced depressive-like behavior through inhibition of toll-like receptor 4 expression via the PI3K/AKT/FoxO1 pathway. J Neuroinflammation 2019 May 8;16(1):95 (PubMed: 31068207) [IF=5.793]

Application: WB    Species: mouse    Sample: hippocampus

Fig. 3| The role of TLR4 in alleviating neuroinflammation of baicalin. Prophylactic treatment of TAK-242 or baicalin inhibited LPS-induced neuroinflammation as well as depressive-like behaviors. d The levels of TLR4 and p-FoxO1 in the hippocampus were measured by western blotting and were quantified and normalized with their respective β-actin or total FoxO1 levels (n = 4). Data were normalized to the control and presented as means ± SEM. #p < 0.05, ##p < 0.01, ###p < 0.005 vs. control; *p < 0.05, **p < 0.01, ***p < 0.005 vs. LPS. TAK, TAK-242

Application: IF/ICC    Species: mouse    Sample: BV2 cells

Fig. 7 |Baicalin regulates LPS-induced TLR4 expression by activating the PI3K/AKT/FoxO1 pathway in BV2 cells.e BV2 cells were stained with a TLR4 antibody and observed with fluorescence microscopy. Green, TLR4; blue: DAPI; scale bar, 50 μm

9). Li S et al. Dental pulp stem cell‐derived exosomes alleviate cerebral ischaemia‐reperfusion injury through suppressing inflammatory response. Cell Prolif 2021 Aug;54(8):e13093. (PubMed: 34231932) [IF=5.753]

10). Wang S et al. Overexpression of Toll-Like Receptor 4 Affects Autophagy, Oxidative Stress, and Inflammatory Responses in Monocytes of Transgenic Sheep. Front Cell Dev Biol 2020 May 5;8:248. (PubMed: 32432106) [IF=5.201]

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