TLR4 Antibody (76B357.1) - Azide Free

TLR4 (Toll-like receptor 4) is a type-1 transmembrane glycoprotein that is a pattern recognition receptor (PRR) belonging to the TLR family (1-3). TLR4 is expressed in many tissues and is most abundantly expressed in the placenta, spleen, and peripheral blood leukocytes (1). Human TLR4 is synthesized as a 839 amino acid (aa) protein containing a signal sequence (1-23 aa), an extracellular domain (ECD) (24-631 aa), a transmembrane domain (632-652 aa), and Toll/interleukin-1 receptor (TIR) cytoplasmic domain (652-839 aa) with a theoretical molecular weight of 95 kDa (3, 4). The ECD contains 21 leucine-rich repeats (LRRs) and has a horseshoe-shaped structure (3, 4). TLR4 requires binding with the co-receptor myeloid differentiation protein 2 (MD2) largely via hydrophilic interactions for proper ligand sensing and signaling (2-4). In general, the TLR family plays a role in activation of innate immunity and responds to a variety of pathogen-associated molecular patterns (PAMPs) (5). TLR4 is specifically responsive to lipopolysaccharide (LPS), which is found on the outer-membrane of most ram-negative bacteria (3-5). Activation of TLR4 requires binding of a ligand, such as LPS to MD2, followed by MD2-LPS complex binding to TLR4, resulting in a partial complex (TLR4-MD2/LPS) (3, 5). To become fully active, two partial complexes must dimerize thereby allowing the TIR domains of TLR4 to bind other adapter molecular and initiate signaling, triggering an inflammatory response and cytokine production (3, 5).

TLR4 signaling occurs through two distinct pathways: The MyD88 (myeloid differentiation primary response gene 88)-dependent pathway and the MyD88-independent (TRIF-dependent, TIR domain-containing adaptor inducing IFN-beta) pathway (3, 5-7). The MyD88-dependent pathway occurs mainly at the plasma membrane and involves the binding of MyD88-adaptor-like (MAL) protein followed by a signaling cascade that results in the activation of transcription factors including nuclear factor-kappaB (NF-kappaB) that promote the secretion of inflammatory molecules and increased phagocytosis (5-7). Conversely, the MyD88-independent pathway occurs after TLR4-MD2 complex internalization in the endosomal compartment. This pathway involves the binding of adapter proteins TRIF and TRIF-related adaptor molecule (TRAM), a signaling activation cascade resulting in IFN regulatory factor 3 (IRF3) translocation into the nucleus, and secretion of interferon-beta (INF-beta) genes and increased phagocytosis (5-7).

Given its expression on immune-related cells and its role in inflammation, TLR4 activation can contribute to various diseases (6-8). For instance, several studies have found that TLR4 activation is associated with neurodegeneration and several central nervous system (CNS) pathologies, including Alzheimer's disease, Parkinson's disease, and Huntington's disease (6, 7). Furthermore, TLR4 mutations have been shown to lead to higher rates of infections and increased susceptibility to sepsis (7-8). One potential therapeutic approach aimed at targeting TLR4 and neuroinflammation is polyphenolic compounds which include flavonoids and phenolic acids and alcohols (8).

Alternative names for TLR4 includes 76B357.1, ARMD10, CD284 antigen, CD284, EC 3.2.2.6, homolog of Drosophila toll, hToll, toll like receptor 4 protein, TOLL, toll-like receptor 4.

References

1. Vaure, C., & Liu, Y. (2014). A comparative review of toll-like receptor 4 expression and functionality in different animal species. Frontiers in immunology. https://doi.org/10.3389/fimmu.2014.00316

2. Park, B. S., & Lee, J. O. (2013). Recognition of lipopolysaccharide pattern by TLR4 complexes. Experimental & molecular medicine. https://doi.org/10.1038/emm.2013.97

3. Krishnan, J., Anwar, M.A., & Choi, S. (2016) TLR4 (Toll-Like Receptor 4). In: Choi S. (eds) Encyclopedia of Signaling Molecules. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-6438-9_592-1

4. Botos, I., Segal, D. M., & Davies, D. R. (2011). The structural biology of Toll-like receptors. Structure. https://doi.org/10.1016/j.str.2011.02.004

5. Lu, Y. C., Yeh, W. C., & Ohashi, P. S. (2008). LPS/TLR4 signal transduction pathway. Cytokine. https://doi.org/10.1016/j.cyto.2008.01.006

6. Leitner, G. R., Wenzel, T. J., Marshall, N., Gates, E. J., & Klegeris, A. (2019). Targeting toll-like receptor 4 to modulate neuroinflammation in central nervous system disorders. Expert opinion on therapeutic targets. https://doi.org/10.1080/14728222.2019.1676416

7. Molteni, M., Gemma, S., & Rossetti, C. (2016). The Role of Toll-Like Receptor 4 in Infectious and Noninfectious Inflammation. Mediators of inflammation. https://doi.org/10.1155/2016/6978936

8. Rahimifard, M., Maqbool, F., Moeini-Nodeh, S., Niaz, K., Abdollahi, M., Braidy, N., Nabavi, S. M., & Nabavi, S. F. (2017). Targeting the TLR4 signaling pathway by polyphenols: A novel therapeutic strategy for neuroinflammation. Ageing research reviews. https://doi.org/10.1016/j.arr.2017.02.004

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

We have publications tested in 9 applications: B/N, FLOW, Flow Cytometry Control, Flow-CS, ICC/IF, IHC-Fr, IHC-P, WB, Western Blot.

Showing Publications 1 - 10 of 30. Show All 30 Publications. Collapse Publications.

Publications using NBP2-27149	Applications	Species
Li L, Kong X, Chen W et al. Guizhi-Shaoyao-Zhimu decoction attenuates rheumatoid arthritis by inhibiting mast cell degranulation Traditional Medicine Research 2023-02-23 (Western Blot, Rat)	Western Blot	Rat
Vobeta EV, Skuljec J, Gudi V et al. Characterisation of microglia during de- and remyelination: Can they promote a repair promoting environment Neurobiology of Disease Details: This publication used the FITC conjugated form of this antibody (Cat# NB100-55951).
Khan HU, Aamir K, Jusuf PR et al Lauric acid ameliorates lipopolysaccharide (LPS)-induced liver inflammation by mediating TLR4/MyD88 pathway in Sprague Dawley (SD) rats Life Sci 2020-11-14 [PMID: 33188836] (WB, Mouse) Details: Citation using the Azide Free version of this antibody.	WB	Mouse
Najjar R Raspberry Polyphenols Target Molecular Pathways of Heart Failure Thesis 2023-01-01
Bi Y, Ding Y, Wu J Et al. Staphylococcus aureus induces mammary gland fibrosis through activating the TLR/NF-kB and TLR/AP-1 signaling pathways in mice Microb Pathog 2020-08-14 [PMID: 32783982] Details: Citation using the Azide Free version of this antibody.
Haricharan S, Brown P. TLR4 has a TP53-dependent dual role in regulating breast cancer cell growth Proc. Natl Acad Sci U. [PMID: 26063617] (B/N, Human) Details: Citation using the Azide Free form of this antibody.	B/N	Human
Degraaf AJ, Zas?ona Z, Bourdonnay E, Peters-Golden M. Prostaglandin E2 reduces Toll-like Receptor 4 Expression in Alveolar Macrophages by Inhibition of Translation. Am. J. Respir. Cell Mol. Biol. 2014-03-06 [PMID: 24601788] (Flow-CS, Rat) Details: This citation used the PE version of this antibody.	Flow-CS	Rat
Abdi J, Mutis T, Garssen J, Redegeld F. Characterization of the Toll-like receptor expression profile in human multiple myeloma cells. PLoS One. 2013-04-08 [PMID: 23593278] Details: Antibodies cited [multiple myeloma (MM) cell lines, human intestine tissue lysate, PBMC, mouse splenocytes] for WB (Figs 2, 3, 4A, 4B, 5, Table 1) and Flow (Intracellular): Figs 4C, 6-8, Table 1: 1. TLR1 (IMG-5012) 2. TLR2 (IMG-416A)3. TLR3 (IMG-315A)4 TL
Semar S, Klotz M, Letiembre M et al. Changes of the enteric nervous system in amyloid-B protein precursor transgenic mice correlate with disease progression. J Alzheimers Dis. 2013-01-01 [PMID: 23531500]
Jazaeri F, Tavangar SM, Ghazi-Khansari M et al. Cirrhosis is associated with development of tolerance to cardiac chronotropic effect of endotoxin in rats. Liver Int. 2013-03-01 [PMID: 23311391] Details: IMG-5031A (clone 76B357.1), IHC (P): Fig 3 (rat atrial tissue). TLR4 staining was observed in myocytes and endothelial cells. LPS upregulated TLR4 expression. Human breast tissue was used as positive control.
McCulloch L, Brown KL, Mabbott NA. Ablation of the cellular prion protein, PrPC, specifically on follicular dendritic cells has no effect on their maturation or function. Immunology. 2013-03-01 [PMID: 23121447] (IHC-Fr, Mouse) Details: IMG-5031A (clone 76B357.1), IHC (F): 6 (mouse spleen).	IHC-Fr	Mouse
Hsiao CC, Kao YH, Huang SC, Chuang JH. Toll-like receptor-4 agonist inhibits motility and invasion of hepatoblastoma HepG2 cells in vitro. Pediatr Blood Cancer. 2013-02-01 [PMID: 22648929] Details: TLR4/CD284 (IMG-5031A). IHC (P): Fig 1 (human hepatoblastoma). TLR4 staining was membranous, cytoplasmic and nuclear. TLR4 staining was greatly reduced in liver tissues obtained after chemotherapy treatment.
Zheng YX, Yang M, Rong TT et al. CD74 and macrophage migration inhibitory factor as therapeutic targets in gastric cancer. World J Gastroenterol. 2012-05-14 [PMID: 22611320] (IHC-P, WB, Human)	IHC-P, WB	Human
Liu Y, Fatheree NY, Mangalat N, Rhoads JM. Lactobacillus reuteri strains reduce incidence and severity of experimental necrotizing enterocolitis via modulation of TLR4 and NF-kB signaling in the intestine. Am J Physiol Gastrointest Liver Physiol. 2012-03-15 [PMID: 22207578]
Janardhanam SB, Prakasam S, Swaminathan VT et al. Differential expression of TLR-2 and TLR-4 in the epithelial cells in oral lichen planus. Arch Oral Biol. 2012-05-01 [PMID: 22119043] Details: IHC (P) of human oral epithelium (Fig 1): 1. TLR2 (IMG-319) 2. TLR4 (IMG-5031A) Note: Both antibodies were used at 0.5/ml. Cutaneous epithelium was used as a positive control for TLR2 and TLR4 expression.
Weile C, Josefsen K, Buschard K. Glucose activation of islets of Langerhans up-regulates Toll-like receptor 5: possible mechanism of protection. Clin Exp Immunol. 2011-11-01 [PMID: 21985371] Details: TLR4/CD284 (IMG-5031A) and GAPDH (IMG-3073). WB: Cultured rat islet Langerhans treated with glucose, Fig 1B. Note: TLR4 protein levels were not increased in the presence of glucose activation, Fig 1B.
Vobeta EV, Skuljec J, Gudi V et al. Characterisation of microglia during de- and remyelination: Can they promote a repair promoting environment Neurobiol Dis 2011-10-06 [PMID: 21971527]
Nowicki M, Kosacka J, Serke H et al. Altered sciatic nerve fiber morphology and endoneural microvessels in mouse models relevant for obesity, peripheral diabetic polyneuropathy, and the metabolic syndrome. J Neurosci Res. 2012-01-01 [PMID: 21919033] Details: TLR4 IMG-5031A (clone 76B357.1) for IHC (cryostat sections): Fig 5 (dissected mouse nerves). Note: the antibody was used at 1:500. Mice were perfused with 4% formaldehyde in 0.1 M PBS prior to dissection.
Brandau S, Jakob M, Hemeda H et al. Tissue-resident mesenchymal stem cells attract peripheral blood neutrophils and enhance their inflammatory activity in response to microbial challenge. J Leukoc Biol. 2010-11-01 [PMID: 20682625] (FLOW, Human) Details: TLR4 antibody clone 76B357.1 cited for Flow (Cell Surface), Fig 2: human primary parotid mesenchymal stem cell cultures.	FLOW	Human
Hafez M, Hayes K, Goldrick M et al. The K5 capsule of Escherichia coli strain Nissle 1917 is important in stimulating expression of Toll-like receptor 5, CD14, MyD88, and TRIF together with the induction of interleukin-8 expression via the mitogen-activated protein kinase pathway in epithelial cells. Infect Immun. 2010-05-01 [PMID: 20145095] Details: IMGENEX products cited: 1. TLR4 (IMG- 5031A) Flow (Cell surface), Fig 3, Caco-2 cells. 2. TLR5 (IMG-663A) Flow (Cell surface), Fig 2, Fig 3, Caco-2 cells.
Johnston MJ, Wang A, Catarino ME et al. Extracts of the rat tapeworm, Hymenolepis diminuta, suppress macrophage activation in vitro and alleviate chemically induced colitis in mice. Infect Immun. 2010-03-01 [PMID: 20028812]
Wu J, Meng Z, Jiang M et al. Toll-like receptor-induced innate immune responses in non-parenchymal liver cells are cell type-specific. Immunology. [PMID: 19922426] Details: Citation using the PE/Cy5 form of this antibody.
Nowicki M, Muller K, Serke H et al. Oxidized low-density lipoprotein (oxLDL)-induced cell death in dorsal root ganglion cell cultures depends not on the lectin-like oxLDL receptor-1 but on the toll-like receptor-4. J Neurosci Res. 2010-02-01 [PMID: 19705455] (ICC/IF, Rat) Details: IF: DRG (dorsal root ganglia) cells from ten day postnatal SPRD male rats, Fig 5B.	ICC/IF	Rat
Worku M, Morris A. Binding of different forms of lipopolysaccharide and gene expression in bovine blood neutrophils. J Dairy Sci 2009-07-01 [PMID: 19528595] Details: Using the PE conjugated version of NBP2-27149, catalog number NBP2-27149PE.
Link A, Selejan S, Maack C et al. Phosphodiesterase 4 inhibition but not beta-adrenergic stimulation suppresses tumor necrosis factor-alpha release in peripheral blood mononuclear cells in septic shock. Crit Care. 2008-01-01 [PMID: 19091080] Details: TLR4/CD284 (IMG-5031C): Flow (cell surface), human CD14+ monocytes derived from healthy donors stimulated with increasing concentrations of LPS (lipopolysaccharide). 2. TLR2/CD282 (IMG-6320C): Flow (cell surface), human CD14+ monocytes derived from healthy donors stimulated with increasing concentrations of SEB (Staphylococcus aureus enterotoxin).
Lunardi C, Bason C, Dolcino M et al. Antiflagellin antibodies recognize the autoantigens Toll-Like Receptor 5 and Pals 1-associated tight junction protein and induce monocytes activation and increased intestinal permeability in Crohn's disease. J Intern Med. 2009-02-01 [PMID: 18796002] Details: WB (IMG-5031A): Fig 1a (293T cells transfected with the human TLR4 gene). TLR4 was detected in the TLR4 293T transfected cells (lane 2, ~88 kDa), but not in the untransfected cells (lane 1). TLR4 was observed as an ~88 kDa band in lane 2. Note: The specificity of the antibody was TLR4 transfected validated in Fig 1 by western blot [compare lanes 1 (untransfected) and 2 (TLR4 transfected)].
Cohen PA, Koski GK, Czerniecki BJ et al. STAT3- and STAT5-dependent pathways competitively regulate the pan-differentiation of CD34pos cells into tumor-competent dendritic cells. Blood. 2008-09-01 [PMID: 18577706] (Flow Cytometry Control, Mouse) Details: Flow (intracellular), mouse bone marrow cells, Fig. 1E: 1. TLR3 FITC (IMG-315C) 2. TLR4 FITC (IMG-417C) 3. TLR7 (IMG-665A) 4. TLR8 FITC (IMG-321C) 5. TLR9 FITC (IMG-305C).	Flow Cytometry Control	Mouse
Yoshizaki A, Iwata Y, Komura K et al. CD19 regulates skin and lung fibrosis via Toll-like receptor signaling in a model of bleomycin-induced scleroderma. Am J Pathol. 2008-06-01 [PMID: 18467694]
Allam JP, Peng WM, Appel T et al. Toll-like receptor 4 ligation enforces tolerogenic properties of oral mucosal Langerhans cells. J Allergy Clin Immunol. 2008-02-01 [PMID: 18036651] (Flow-CS, Human) Details: Flow (cell surface): Oral mucosa from the vestibular region were obtained from non-tumor and noninflamed human patients, Fig 1C.	Flow-CS	Human
Zanoni G, Navone R, Lunardi C et al. In Celiac Disease, a Subset of Autoantibodies Against Transglutaminase Binds Toll-Like Receptor 4 and Induces Activation of Monocytes PLoS Med 2006-09-01 [PMID: 16984219]
Show All 30 Publications. Collapse Publications.

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