Recombinant Human TLR4/MD2 Complex Protein Summary
Product Specifications
| Human TLR4 (Glu24-Lys631) Accession # O00206.2 | S | 10-His tag |
| Human MD-2 (Glu17-Asn160) Accession # Q9Y6Y9.1 | IEGRGGGSGGGSGGGS | 10-His tag |
| N-terminus | C-terminus | |
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
3146-TM
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
3146-TM/CF
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile PBS. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Reconstitution Calculator
Background: TLR4/MD2 Complex
TLR4 is a 100 kDa type I transmembrane glycoprotein that belongs to the mammalian Toll-Like Receptor family of pathogen pattern recognition molecules. MD‑2, also known as ESOP-1, is a 25 kDa secreted protein that is required for TLR4‑mediated responses to bacterial lipopolysaccharide (LPS) (1 - 3). The human TLR4 cDNA encodes an 839 amino acid (aa) precursor that contains a 23 aa signal sequence, a 608 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 187 aa cytoplasmic domain. TLR4 contains 21 leucine rich repeats in its ECD and one cytoplasmic Toll/IL-1 receptor (TIR) domain (4). The ECD of human TLR4 shares approximately 25% aa sequence identity with other TLRs and 60% - 74% aa sequence identity with bovine, equine, feline, mouse, rat, and porcine TLR4. The human MD‑2 cDNA encodes a 160 aa precursor with an 18 aa signal sequence (5). Human MD‑2 shares 20% aa sequence identity with MD‑1 and 62% - 64% aa sequence identity with bovine, mouse, and rat MD‑2. MD‑2 associates with TLR4 on monocytes, macrophages, dendritic cells, and B cells (5 - 7). MD2 expression is required for cell surface localization of TLR4 and for optimal LPS‑induced TLR4 signaling (7, 8). MD‑2 also forms soluble disulfide-linked homo-oligomers which can interact with TLR4 (6). Through a domain separate from its TLR4-binding domain, MD‑2 extracts LPS from circulating CD14‑LPS complexes and carries the LPS into a ternary complex with TLR4 (9 - 11). The interaction of MD‑2/LPS with TLR4 induces receptor oligomerization and the triggering of an inflammatory response (12). Increased levels of plasma MD‑2 in septic shock patients sensitizes MD‑2 non-expressing epithelial cells to LPS and promotes widespread tissue inflammation (13).
- Gangloff, M. and N.J. Gay (2004) Trends Biochem. Sci. 29:294.
- Palsson-McDermott, E.M. and L.A. O’Neill (2004) Immunology 113:153.
- Miyake, K. (2004) Semin. Immunol. 16:11.
- Medzhitov, R. et al. (1997) Nature 388:394.
- Shimazu, R. et al. (1999) J. Exp. Med. 189:1777.
- Visintin, A. et al. (2001) Proc. Natl. Acad. Sci. USA 98:12156.
- Akashi, S. et al. (2000) J. Immunol. 164:3471.
- Nagai, Y. et al. (2002) Nat. Immunol. 3:667.
- Re, F. and J.L. Strominger (2003) J. Immunol. 171:5272.
- Kennedy, M.N. et al. (2004) J. Biol. Chem. 279:34698.
- Gioannini, T.L. et al. (2004) Proc. Natl. Acad. Sci. USA 101:4186.
- Saitoh, S. et al. (2004) J. Endotoxin Res. 10:257.
- Pugin, J. et al. (2004) Blood 104:4071.
Citations for Recombinant Human TLR4/MD2 Complex Protein
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
23
Citations: Showing 1 - 10
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The bacterial lysate OM-85 engages Toll-like receptors 2 and 4 triggering an immunomodulatory gene signature in human myeloid cells
Authors: Khameneh, HJ;Bolis, M;Ventura, PMO;Cassanmagnago, GA;Fischer, BA;Zenobi, A;Guerra, J;Buzzago, I;Bernasconi, M;Zaman, GJR;Rinaldi, A;Moro, SG;Sallusto, F;Baulier, E;Pasquali, C;Guarda, G;
Mucosal immunology
Species: N/A
Sample Types: Small Molecule
Applications: Bioassay -
Characterization of the Antimicrobial Activities of Trichoplusia ni Cecropin A as a High-Potency Therapeutic against Colistin-Resistant Escherichia coli
Authors: Lee, H;Kim, B;Kim, M;Yoo, S;Lee, J;Hwang, E;Kim, Y;
Pharmaceutics
Species: Human
Sample Types: Recombinant Protein
Applications: Surface Plasmon Resonance -
DNA-mediated proteolysis by neutrophil elastase enhances binding activities of the HMGB1 protein
Authors: X Wang, M Mayorga-Fl, KG Bien, AO Bailey, J Iwahara
The Journal of Biological Chemistry, 2022-10-08;0(0):102577.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Ginsenoside Ro, an oleanolic saponin of Panax ginseng, exerts anti-inflammatory effect by direct inhibiting toll like receptor 4 signaling pathway
Authors: HL Xu, GH Chen, YT Wu, LP Xie, ZB Tan, B Liu, HJ Fan, HM Chen, GQ Huang, M Liu, YC Zhou
Journal of Ginseng Research, 2021-06-05;46(1):156-166.
Species: Human
Sample Types: Recombinant Protein
Applications: Surface Plasmon Resonance -
Venestatin from parasitic helminths interferes with receptor for advanced glycation end products (RAGE)-mediated immune responses to promote larval migration
Authors: D Tsubokawa, T Kikuchi, JM Lee, T Kusakabe, Y Yamamoto, H Maruyama
PloS Pathogens, 2021-06-03;17(6):e1009649.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Toll-like receptor 4-mediated inflammation triggered by extracellular IFI16 is enhanced by lipopolysaccharide binding
Authors: A Iannucci, V Caneparo, S Raviola, I Debernardi, D Colangelo, R Miggiano, G Griffante, S Landolfo, M Gariglio, M De Andrea
PLoS Pathog., 2020-09-09;16(9):e1008811.
Species: Human
Sample Types: Protein
Applications: Surface Plasmon Resonance -
Towards automation in biologics production via Raman micro-spectroscopy, laser-induced forward cell transfer and surface-enhanced Raman spectroscopy
Authors: E Jaeckle, E Brauchle, N Nottrodt, M Wehner, R Lensing, A Gillner, K Schenke-La, M Bach, A Burger-Ken
J. Biotechnol., 2020-09-06;0(0):.
Species: Hamster
Sample Types: Cell Culture Supernates
Applications: Western Blot -
HIV-1 Tat - TLR4/MD2 interaction drives the expression of IDO-1 in monocytes derived dendritic cells through NF-kappaB dependent pathway
Authors: E Bahraoui, M Serrero, R Planès
Sci Rep, 2020-05-18;10(1):8177.
Species: Human
Sample Types: Recombinant Protein, Whole Cells
Applications: Bioassay, ELISA Capture -
Antimicrobial Peptides Derived from the Immune Defense Protein CAP37 Inhibit TLR4 Activation by S100A9
Authors: A Kasus-Jaco, CA Land, AJ Stock, JL Washburn, HA Pereira
Invest. Ophthalmol. Vis. Sci., 2020-04-09;61(4):16.
Species: Human
Sample Types: Protein
Applications: ELISA -
Autoinhibitory regulation of S100A8/S100A9 alarmin activity locally restricts sterile inflammation
Authors: T Vogl, A Stratis, V Wixler, T Völler, S Thurainaya, SK Jorch, S Zenker, A Dreiling, D Chakrabort, M Fröhling, P Paruzel, C Wehmeyer, S Hermann, O Papantonop, C Geyer, K Loser, M Schäfers, S Ludwig, M Stoll, T Leanderson, JL Schultze, S König, T Pap, J Roth
J. Clin. Invest., 2018-04-03;0(0):.
Species: Human
Sample Types: Peptide
Applications: Bioassay -
Folic acid derived-P5779 mimetics regulate DAMP-mediated inflammation through disruption of HMGB1:TLR4:MD-2 axes
Authors: S Sun, M He, Y Wang, H Yang, Y Al-Abed
PLoS ONE, 2018-02-15;13(2):e0193028.
Applications: Bioassay -
Rational Design of a New Class of Toll-Like Receptor 4 (TLR4) Tryptamine Related Agonists by Means of the Structure- and Ligand-Based Virtual Screening for Vaccine Adjuvant Discovery
Authors: J Honegr, R Dolezal, D Malinak, M Benkova, O Soukup, JSFD Almeida, TCC Franca, K Kuca, R Prymula
Molecules, 2018-01-04;23(1):.
Applications: Bioassay -
Toll-like Receptor 2 Antagonists Identified through Virtual Screening and Experimental Validation
Authors: P Durai, HJ Shin, A Achek, HK Kwon, RG Govindaraj, S Panneersel, D Yesudhas, J Choi, KT No, S Choi
FEBS J., 2017-07-02;0(0):.
Applications: Surface Plasmon Resonance -
Extracellular Histones Induce Chemokine Production in Whole Blood Ex Vivo and Leukocyte Recruitment In Vivo.
Authors: Westman J, Papareddy P, Dahlgren M, Chakrakodi B, Norrby-Teglund A, Smeds E, Linder A, Morgelin M, Johansson-Lindbom B, Egesten A, Herwald H
PLoS Pathog, 2015-12-08;11(12):e1005319.
Applications: Surface Plasmon Resonance -
Enhancement of tumor-specific T cell-mediated immunity in dendritic cell-based vaccines by Mycobacterium tuberculosis heat shock protein X.
Authors: Jung I, Shin S, Lee M, Kang T, Han H, Lee S, Kim W, Kim H, Park W, Kim H, Yun C, Lee E, Wu T, Park Y
J Immunol, 2014-07-02;193(3):1233-45.
Applications: Bioassay -
Selective antibody intervention of Toll-like receptor 4 activation through Fc gamma receptor tethering.
Authors: Shang L, Daubeuf B, Triantafilou M, Olden R, Depis F, Raby A, Herren S, Dos Santos A, Malinge P, Dunn-Siegrist I, Benmkaddem S, Geinoz A, Magistrelli G, Rousseau F, Buatois V, Salgado-Pires S, Reith W, Monteiro R, Pugin J, Leger O, Ferlin W, Kosco-Vilbois M, Triantafilou K, Elson G
J Biol Chem, 2014-04-15;289(22):15309-18.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Zhankuic acid A isolated from Taiwanofungus camphoratus is a novel selective TLR4/MD-2 antagonist with anti-inflammatory properties.
Authors: Chen, Yu-Fon, Shiau, Ai-Li, Wang, Sheng-Hu, Yang, Jai-Sing, Chang, Sue-Joan, Wu, Chao-Lia, Wu, Tian-Shu
J Immunol, 2014-02-14;192(6):2778-86.
Species: Bacteria - Taiwanofungus camphoratus
Sample Types: Protein
Applications: Bioassay -
Maximizing the potency of an anti-TLR4 monoclonal antibody by exploiting proximity to Fcgamma receptors.
Authors: Loyau J, Malinge P, Daubeuf B, Shang L, Elson G, Kosco-Vilbois M, Fischer N, Rousseau F
MAbs, 2014-01-01;6(6):1621-30.
Species: Human
Sample Types: Antibody
Applications: Surface Plasmon Resonance -
Fetuin-A acts as an endogenous ligand of TLR4 to promote lipid-induced insulin resistance.
Nat Med, 2012-07-29;18(0):1279.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Sialyl residues modulate LPS-mediated signaling through the Toll-like receptor 4 complex.
Authors: Feng C, Stamatos NM, Dragan AI, Medvedev A, Whitford M, Zhang L, Song C, Rallabhandi P, Cole L, Nhu QM, Vogel SN, Geddes CD, Cross AS
PLoS ONE, 2012-04-09;7(4):e32359.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding.
Authors: Sakaguchi M, Murata H, Yamamoto K, Ono T, Sakaguchi Y, Motoyama A, Hibino T, Kataoka K, Huh NH
PLoS ONE, 2011-08-01;6(8):e23132.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Non-acylated Mycobacterium bovis glycoprotein MPB83 binds to TLR1/2 and stimulates production of matrix metalloproteinase 9.
Authors: Chambers MA, Whelan AO, Spallek R
Biochem. Biophys. Res. Commun., 2010-08-26;400(3):403-8.
Species: Bacteria
Sample Types: Recombinant Protein
Applications: Surface Plasmon Resonance -
Soluble forms of Toll-like receptor 4 are present in human saliva and modulate tumour necrosis factor-alpha secretion by macrophage-like cells.
Authors: Zunt SL, Burton LV, Goldblatt LI, Dobbins EE, Srinivasan M
Clin. Exp. Immunol., 2009-03-09;156(2):285-93.
Applications: Western Blot
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Reason for Rating: we use this protein complex to see protein-protein interaction with allergic proteins. TLR4/MD2 complex detects both TLR4 and MD2 antibodies very well at a minimum of 1:10 folds in direct ELISA.
