Recombinant Human TLR4/MD2 Complex Protein

Name: Recombinant Human TLR4/MD2 Complex Protein
Brand: R&D Systems
SKU: 3146-TM
Rating: 4 (1 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # 3146-TM

Citations (21)

(1)

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Setting the standard in quality research reagents

Key Product Details

Source:	NS0
Accession #:	O00206.2 (TLR4) & Q9Y6Y9.1 (MD-2)
Applications:	Bioactivity

View Full Technical Details for Recombinant Human TLR4/MD2 Complex Protein

Product Specifications for Recombinant Human TLR4/MD2 Complex Protein

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

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.

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

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.

Formulation, Preparation and Storage

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.

3146-TM

Carrier

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

Carrier Free

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

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).

Item	Value
References	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.
Long Name	Toll-like Receptor 4
Entrez Gene IDs	7099 (Human)
Alternate Names	TLR4/MD-2 Complex, TLR4/MD2 Complex

Citations for Recombinant Human TLR4/MD2 Complex Protein (21)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human TLR4/MD2 Complex Protein include:

Species: Human
Sample Types: Recombinant Protein
Applications: Surface Plasmon Resonance

HL Xu et al. (2022), Ginsenoside Ro, an oleanolic saponin of Panax ginseng, exerts anti-inflammatory effect by direct inhibiting toll like receptor 4 signaling pathway Journal of Ginseng Research, 46(1):156-166.
PMID: 35058732
Species: Human
Sample Types: Protein
Applications: Bioassay

X Wang et al. (2022), DNA-mediated proteolysis by neutrophil elastase enhances binding activities of the HMGB1 protein The Journal of Biological Chemistry, 0(0):102577.
PMID: 36220391
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay

D Tsubokawa et al. (2021), Venestatin from parasitic helminths interferes with receptor for advanced glycation end products (RAGE)-mediated immune responses to promote larval migration PloS Pathogens, 17(6):e1009649.
PMID: 34081755
Species: Human
Sample Types: Protein
Applications: ELISA

A Kasus-Jaco et al. (2020), Antimicrobial Peptides Derived from the Immune Defense Protein CAP37 Inhibit TLR4 Activation by S100A9 Invest. Ophthalmol. Vis. Sci., 61(4):16.
PMID: 32298435
Species: Human
Sample Types: Protein
Applications: Surface Plasmon Resonance

A Iannucci et al. (2020), Toll-like receptor 4-mediated inflammation triggered by extracellular IFI16 is enhanced by lipopolysaccharide binding PLoS Pathog., 16(9):e1008811.
PMID: 32903274
Species: Human
Sample Types: Recombinant Protein
Applications: ELISA Capture

E Bahraoui et al. (2020), HIV-1 Tat - TLR4/MD2 interaction drives the expression of IDO-1 in monocytes derived dendritic cells through NF-kappaB dependent pathway Sci Rep, 10(1):8177.
PMID: 32424165
Species: Hamster
Sample Types: Cell Culture Supernates
Applications: Western Blot

E Jaeckle et al. (2020), Towards automation in biologics production via Raman micro-spectroscopy, laser-induced forward cell transfer and surface-enhanced Raman spectroscopy J. Biotechnol., 0(0).
PMID: 32898625
Sample Types: Complex Sample Type
Applications: Bioassay

S Sun et al. (2018), Folic acid derived-P5779 mimetics regulate DAMP-mediated inflammation through disruption of HMGB1:TLR4:MD-2 axes PLoS ONE, 13(2):e0193028.
PMID: 29447234
Species: Human
Sample Types: Peptide
Applications: Bioassay

T Vogl et al. (2018), Autoinhibitory regulation of S100A8/S100A9 alarmin activity locally restricts sterile inflammation J. Clin. Invest., 0(0).
PMID: 29611822
Applications: Bioassay

J Honegr et al. (2018), 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 Molecules, 23(1).
PMID: 29300367
Species: N/A
Sample Types: Recombinant Protein
Applications: Surface Plasmon Resonance

P Durai et al. (2017), Toll-like Receptor 2 Antagonists Identified through Virtual Screening and Experimental Validation FEBS J., 0(0).
PMID: 28570013
Species: N/A
Sample Types: Peptide
Applications: Surface Plasmon Resonance

Westman J et al. (2015), Extracellular Histones Induce Chemokine Production in Whole Blood Ex Vivo and Leukocyte Recruitment In Vivo. PLoS Pathog, 11(12):e1005319.
PMID: 26646682
Chen et al. (2014), Zhankuic acid A isolated from Taiwanofungus camphoratus is a novel selective TLR4/MD-2 antagonist with anti-inflammatory properties. J Immunol, 192(6):2778-86.
PMID: 24532584
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Shang L et al. (2014), Selective antibody intervention of Toll-like receptor 4 activation through Fc gamma receptor tethering. J Biol Chem, 289(22):15309-18.
PMID: 24737331
Species: N/A
Sample Types: Protein
Applications: Bioassay

Jung I et al. (2014), Enhancement of tumor-specific T cell-mediated immunity in dendritic cell-based vaccines by Mycobacterium tuberculosis heat shock protein X. J Immunol, 193(3):1233-45.
PMID: 24990079
Species: Human
Sample Types: Antibody
Applications: Surface Plasmon Resonance

Loyau J et al. (2014), Maximizing the potency of an anti-TLR4 monoclonal antibody by exploiting proximity to Fcgamma receptors. MAbs, 6(6):1621-30.
PMID: 25484053
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

(2012), Fetuin-A acts as an endogenous ligand of TLR4 to promote lipid-induced insulin resistance. Nat Med, 18(0):1279.
PMID: 22842477
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Feng C et al. (2012), Sialyl residues modulate LPS-mediated signaling through the Toll-like receptor 4 complex. PLoS ONE, 7(4):e32359.
PMID: 22496731
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Sakaguchi M et al. (2011), TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. PLoS ONE, 6(8):e23132.
PMID: 21829704
Species: Bacteria
Sample Types: Recombinant Protein
Applications: Surface Plasmon Resonance

Chambers MA et al. (2010), Non-acylated Mycobacterium bovis glycoprotein MPB83 binds to TLR1/2 and stimulates production of matrix metalloproteinase 9. Biochem. Biophys. Res. Commun., 400(3):403-8.
PMID: 20800577
Species: N/A
Sample Types: N/A
Applications: Western Blot

Zunt SL et al. (2009), Soluble forms of Toll-like receptor 4 are present in human saliva and modulate tumour necrosis factor-alpha secretion by macrophage-like cells. Clin. Exp. Immunol., 156(2):285-93.
PMID: 19292767

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Customer Reviews for Recombinant Human TLR4/MD2 Complex Protein (1)

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Recombinant Human TLR4/MD2 Complex Protein

Name: Anonymous

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.

Verified Customer | Posted 03/29/2021

Recombinant Human TLR4/MD2 Complex Protein

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Product Specifications for Recombinant Human TLR4/MD2 Complex Protein

Formulation, Preparation and Storage

Carrier Free

3146-TM

Carrier

3146-TM/CF

Carrier Free

Reconstitution Calculator

Background: TLR4/MD2 Complex

Citations for Recombinant Human TLR4/MD2 Complex Protein (21)

Customer Reviews for Recombinant Human TLR4/MD2 Complex Protein (1)

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Recombinant Human TLR4/MD2 Complex Protein

Product Documents for Recombinant Human TLR4/MD2 Complex Protein

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