TLR4 Antibody (76B357.1) [Alexa Fluor® 647]

Product Discontinued

TLR4 Antibody (76B357.1) [Alexa Fluor® 647] Summary

• Immunogen: This antibody was developed against a portion of amino acids 100-200 of human TLR4 (NP_612564).
• Specificity: The amino acid sequence used has 100% identity in human, chimp, baboon, chinese hamster, and rat; 93% identity in bovine and pig; 85% identity in sheep and 78% identity in cat, horse and mouse.
• Predicted Species: Porcine (93%), Baboon (100%), Bovine (93%), Chinese Hamster (100%). Backed by our 100% Guarantee.
• Isotype: IgG2b Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: TLR4
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1ul/1 million cells
  • In vivo assay
• Application Notes: Flow (Intracellular) use reported in literature (See Cohen et al, 2008); Immunofluorescence (See Nowicki et al, 2009); IHC-F (Nowicki et al, 2012); Flow (cell surface) use reported in scientific literature (PMID 23796194) Use in In vivo reported in scientific literature (PMID:33498871)
• Agonist:
  • LPS from E. Coli, TLR4 ligand NBP2-25295
• Antagonist:
  • TLR4 Inhibitor Peptide Set NBP2-26244
  • TIRAP (TLR2 and TLR4) Inhibitor Peptide Set NBP2-29331

Reactivity Notes

Ground squirrel reactivity reported by a customer review. Predicted cross-reactivity based on sequence identity: Chimp (100%), Baboon (100%), Chinese Hamster (100%), Bovine (93%), Pig (93%). Mammal reactivity reported in scientific literature (PMID: 25130694)

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: 50mM Sodium Borate
• Preservative: 0.05% Sodium Azide
• Purity: Protein G purified

Notes

Alexa Fluor (R) products are provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment; (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.

Alternate Names for TLR4 Antibody (76B357.1) [Alexa Fluor® 647]

• ARMD10
• CD_antigen: CD284
• CD284 antigen
• CD284
• EC 3.2.2.6
• EC:3.2.2.6
• homolog of Drosophila toll
• hToll
• TLR4
• TLR-4
• toll like receptor 4 protein
• TOLL
• toll-like receptor 4

Background

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

Limitations

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.

Publications for TLR4 Antibody (NBP2-24773)(16)

Publications using NBP2-24773

• Muñoz-Caro T, Gibson AJ, Conejeros I, et al. The Role of TLR2 and TLR4 in Recognition and Uptake of the Apicomplexan Parasite Eimeria bovis and Their Effects on NET Formation Pathogens (Basel, Switzerland) 2021-01-24 [PMID: 33498871] (In Vivo, Mouse)
• Lacave-Lapalun JV, Benderitter M, Linard C Flagellin and LPS each restores rat lymphocyte populations after colorectal irradiation J Leukoc Biol. 2014-06-01 [PMID: 24532644] (Flow-CS, Rat)
  
  Details:
  Citation using the Azide Free format of this antibody.
• Verma R, Jung JH, Kim JY 1,25-Dihydroxyvitamin D3 up-regulates TLR10 while down-regulating TLR2, 4, and 5 in human monocyte THP-1. J Steroid Biochem Mol Biol. 2014-05-01 [PMID: 24373795] (Human)
  
  Details:
  Citation using the PE version of this antibody.
• 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
• 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]
  
  Details:
  IMG-5031A (clone 76B357.1), IHC (F): 6 (mouse spleen).
• 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]
• 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.
• 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.
• 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.
• 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]
  
  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).
• 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]
  
  Details:
  Flow (cell surface): Oral mucosa from the vestibular region were obtained from non-tumor and noninflamed human patients, Fig 1C.
• 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]

Bioinformatics

• Gene Symbol: TLR4