Recombinant Human TLR4/MD2 Complex Protein

Carrier Free

Catalog # Availability Size / Price Qty
3146-TM-050/CF

With Carrier

Catalog # Availability Size / Price Qty
3146-TM-050
R&D Systems Recombinant Proteins and Enzymes
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Product Details
Citations (22)
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Reviews (1)

Recombinant Human TLR4/MD2 Complex Protein Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to inhibit LPS-induced TNF-alpha secretion by PMA-differentiated U937 human histiocytic lymphoma cells. 20 µg/mL of recombinant human TLR4/MD-2 will inhibit >60% of the TNF-alpha secretion induced by LPS.
Source
Mouse myeloma cell line, NS0-derived human TLR4/MD2 Complex protein
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
N-terminal Sequence
Analysis
Glu24 (TLR4) & Glu17 (MD-2)
Predicted Molecular Mass
70.6 kDa (TLR4), 19.2 kDa (MD-2)
SDS-PAGE
95 kDa and 34 kDa, reducing conditions

Product Datasheets

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3146-TM (with carrier)

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3146-TM/CF (carrier free)

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

References
  1. Gangloff, M. and N.J. Gay (2004) Trends Biochem. Sci. 29:294.
  2. Palsson-McDermott, E.M. and L.A. O’Neill (2004) Immunology 113:153.
  3. Miyake, K. (2004) Semin. Immunol. 16:11.
  4. Medzhitov, R. et al. (1997) Nature 388:394.
  5. Shimazu, R. et al. (1999) J. Exp. Med. 189:1777.
  6. Visintin, A. et al. (2001) Proc. Natl. Acad. Sci. USA 98:12156.
  7. Akashi, S. et al. (2000) J. Immunol. 164:3471.
  8. Nagai, Y. et al. (2002) Nat. Immunol. 3:667.
  9. Re, F. and J.L. Strominger (2003) J. Immunol. 171:5272.
  10. Kennedy, M.N. et al. (2004) J. Biol. Chem. 279:34698.
  11. Gioannini, T.L. et al. (2004) Proc. Natl. Acad. Sci. USA 101:4186.
  12. Saitoh, S. et al. (2004) J. Endotoxin Res. 10:257.
  13. Pugin, J. et al. (2004) Blood 104:4071.
Long Name
Toll-like Receptor 4
Entrez Gene IDs
7099 (Human)
Alternate Names
ARMD10;CD284;ESOP-1;lipopolysaccharide response;Ly-96;Lymphocyte antigen 96;myeloid differentiation factor-2;myeloid differentiation protein-2;Protein MD-2;TLR-4;TOLL;Toll-like receptor 4; TLR4/MD2 Complex; TLR4/MD-2 Complex

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.

22 Citations: Showing 1 - 10
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  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. 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
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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
  17. 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
  18. 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
  19. 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
  20. 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
  21. 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
  22. 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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Recombinant Human TLR4/MD2 Complex Protein
By Anonymous on 03/29/2021
Application: Binding assay/Protein-protein interaction
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.