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Human TLR4 Antibody

Catalog #: AF1478
35 Citations Datasheet / COA / SDS
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AF1478
AF1478-SP
TLR4 in Human Prostate.
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Citations (35)
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Human TLR4 Antibody Summary

Species Reactivity
Human
Specificity
Detects human TLR4 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant mouse TLR4 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human TLR4
Glu24-Lys631
Accession # O00206
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Flow Cytometry
2.5 µg/106 cells
Human peripheral blood monocytes
Immunohistochemistry
5-15 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
5-15 µg/mL
See below
Neutralization
Measured by its ability to neutralize LPS-induced IL‑8 secretion in the HEK293 human embryonic kidney cell line co-transfected with human TLR4 and MD-2. The Neutralization Dose (ND50) is typically 1.5-7.5 µg/mL in the presence of 75 ng/mL Lipopolysacharide (LPS).

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunohistochemistry TLR4 antibody in Human Prostate by Immunohistochemistry (IHC-P). View Larger

TLR4 in Human Prostate. TLR4 was detected in immersion fixed paraffin-embedded sections of human prostate using Goat Anti-Human TLR4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1478) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Immunocytochemistry TLR4 antibody in Human PBMCs by Immunocytochemistry (ICC). View Larger

TLR4 in Human PBMCs. TLR4 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human TLR4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1478) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Neutralization IL‑8 Secretion Induced by LPS and Neutralization by Human TLR4 Antibody. View Larger

IL‑8 Secretion Induced by LPS and Neutralization by Human TLR4 Antibody. Lipopolysacharide (LPS) stimulates IL-8 secretion in the HEK293 human embryonic kidney cell line co-transfected with human TLR4 and MD-2, in a dose-dependent manner (orange line), as measured by the Human CXCL8/IL-8 Quantikine ELISA Kit (Catalog # D8000C). IL-8 secretion elicited by LPS (75 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human TLR4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1478). The ND50 is typically 1.5-7.5 µg/mL.

Immunocytochemistry/ Immunofluorescence Detection of Human TLR4 by Immunocytochemistry/ Immunofluorescence View Larger

Detection of Human TLR4 by Immunocytochemistry/ Immunofluorescence Gene expression of biomarkers of injury and immunostaining of TLR4 in patient auricles.(A-D) TLR4 and NOX4 are activated resulting in elevated TNF-alpha in the auricles. Auricles obtained during CABG surgery presented higher expression of TLR4 (P<0.03), BNP (P<0.05), NOX4 (P<0.03), and TNF-alpha (P = 0.135) in reduced versus ‘preserved EF’. (E) TLR2 expression was similar in both groups. (F1-F2) Representative photographs show double-immunostaining of Troponin I and TLR4 in ‘preserved EF’ auricle. (F3-F4) Representative photographs show double-immunostaining of troponin I and TLR4 in the ‘reduced EF’ auricle. TLR4 staining revealed an apparent upregulation in all ‘reduced EF’ patients examined compared to ‘preserved EF’ patients' tissue. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26030867), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunocytochemistry/ Immunofluorescence Detection of Human TLR4 by Immunocytochemistry/ Immunofluorescence View Larger

Detection of Human TLR4 by Immunocytochemistry/ Immunofluorescence Gene expression of biomarkers of injury and immunostaining of TLR4 in patient auricles.(A-D) TLR4 and NOX4 are activated resulting in elevated TNF-alpha in the auricles. Auricles obtained during CABG surgery presented higher expression of TLR4 (P<0.03), BNP (P<0.05), NOX4 (P<0.03), and TNF-alpha (P = 0.135) in reduced versus ‘preserved EF’. (E) TLR2 expression was similar in both groups. (F1-F2) Representative photographs show double-immunostaining of Troponin I and TLR4 in ‘preserved EF’ auricle. (F3-F4) Representative photographs show double-immunostaining of troponin I and TLR4 in the ‘reduced EF’ auricle. TLR4 staining revealed an apparent upregulation in all ‘reduced EF’ patients examined compared to ‘preserved EF’ patients' tissue. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26030867), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: TLR4

TLR4 is a 100 kDa type I transmembrane glycoprotein that belongs to the mammalian Toll-Like Receptor family of pathogen pattern recognition molecules. In the literature molecular weights correspondent to 110 kDa and 130 kDa were reported for TLR4 (1). MD-2, also known as ESOP-1, is a 25 kDa secreted protein that is required for TLR4-mediated responses to bacterial lipopolysaccharide (LPS) (1‑4). 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 (5). 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 (6‑8). MD-2 expression is required for cell surface localization of TLR4 and for optimal LPS-induced TLR4 signaling (8, 9). MD-2 also forms soluble disulfide-linked homo-oligomers which can interact with TLR4 (7). 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 (10‑12). The interaction of MD-2/LPS with TLR4 induces receptor oligomerization and the triggering of an inflammatory response (13). Increased levels of plasma MD-2 in septic shock patients sensitizes MD-2 non-expressing epithelial cells to LPS and promotes widespread tissue inflammation (14).

References
  1. Ohnishi, T. et al. (2003) Clin. Diagn. Lab. Immouol. 10:405.
  2. Gangloff, M. and N.J. Gay (2004) Trends Biochem. Sci. 29:294.
  3. Palsson-McDermott, E.M. and L.A. O’Neill (2004) Immunology 113:153.
  4. Miyake, K. (2004) Semin. Immunol. 16:11.
  5. Medzhitov, R. et al. (1997) Nature 388:394.
  6. Shimazu, R. et al. (1999) J. Exp. Med. 189:1777.
  7. Visintin, A. et al. (2001) Proc. Natl. Acad. Sci. USA 98:12156.
  8. Akashi, S. et al. (2000) J. Immunol. 164:3471.
  9. Nagai, Y. et al. (2002) Nat. Immunol. 3:667.
  10. Re, F. and J.L. Strominger (2003) J. Immunol. 171:5272.
  11. Kennedy, M.N. et al. (2004) J. Biol. Chem. 279:34698.
  12. Gioannini, T.L. et al. (2004) Proc. Natl. Acad. Sci. USA 101:4186.
  13. Saitoh, S. et al. (2004) J. Endotoxin Res. 10:257.
  14. Pugin, J. et al. (2004) Blood 104:4071.
Long Name
Toll-like Receptor 4
Entrez Gene IDs
7099 (Human); 21898 (Mouse); 29260 (Rat)
Alternate Names
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

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Citations for Human TLR4 Antibody

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.

35 Citations: Showing 1 - 10
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  1. PAUF Induces Migration of Human Pancreatic Cancer Cells Exclusively via the TLR4/MyD88/NF-kappa B Signaling Pathway
    Authors: Youn SE, Jiang F, Won HY et al.
    International journal of molecular sciences
  2. CD200 restrains macrophage attack on oligodendrocyte precursors via toll-like receptor 4 downregulation.
    Authors: Hayakawa K, Pham LDD, Seo JH et al.
    J Cereb Blood Flow Metab.
  3. Probiotic effect on Helicobacter�pylori attachment and inhibition of inflammation in human gastric epithelial cells
    Authors: Hanyi Song, Long Zhou, Dongyan Liu, Lihui Ge, Yan Li
    Experimental and Therapeutic Medicine
  4. CCN1 is an opsonin for bacterial clearance and a direct activator of Toll-like receptor signaling
    Authors: JI Jun, LF Lau
    Nat Commun, 2020-03-06;11(1):1242.
  5. Hyaluronic acid restored protein permeability across injured human lung microvascular endothelial cells
    Authors: Shinji Sugita, Yoshifumi Naito, Li Zhou, Hongli He, Qi Hao, Atsuhiro Sakamoto et al.
    FASEB BioAdvances
  6. MicroLet-7b Regulates Neutrophil Function and Dampens Neutrophilic Inflammation by Suppressing the Canonical TLR4/NF-kappa B Pathway
    Authors: Binzhen Chen, Jia Han, Shaoheng Chen, Rufeng Xie, Jie Yang, Tongming Zhou et al.
    Frontiers in Immunology
  7. The effect of Gestational Diabetes Mellitus on the fetal compartment
    Authors: TPB De Luccia, E Ono, R Menon, AU Borbely, R Mattar, L Richardson, ALM da Silva, RM Botelho, MLTLF da Rocha, S Daher
    Journal of reproductive immunology, 2021-03-24;145(0):103314.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  8. Alpha1-antitrypsin protects lung cancer cells from staurosporine-induced apoptosis: the role of bacterial lipopolysaccharide
    Authors: N Schwarz, S Tumpara, S Wrenger, E Ercetin, J Hamacher, T Welte, S Janciauski
    Sci Rep, 2020-06-12;10(1):9563.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  9. Distinct expression patterns of TLR transcripts in human oocytes and granulosa cells from primordial and primary follicles
    Authors: EH Ernst, M Amoushahi, AS Sørensen, TW Kragstrup, E Ernst, K Lykke-Hart
    J. Reprod. Immunol., 2020-04-16;140(0):103125.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  10. Role of TLR4 Receptor Complex in the Regulation of the Innate Immune Response by Fibronectin
    Authors: M Zheng, A Ambesi, P J McKeown-
    Cells, 2020-01-15;9(1):.
    Applications: Neutralizing
  11. Inhibition of osteoclastogenesis by opsonized Porphyromonas gingivalis
    Authors: Subramanya N. Pandruvada, Jeffrey L. Ebersole, Sarandeep S. Huja
    FASEB BioAdvances
  12. sp2-Iminosugar glycolipids as inhibitors of lipopolysaccharide-mediated human dendritic cell activation in vitro and of acute inflammation in mice in vivo
    Authors: Evelyne Schaeffer, Elena M. Sánchez-Fernández, Rita Gonçalves-Pereira, Vincent Flacher, Delphine Lamon, Monique Duval et al.
    European Journal of Medicinal Chemistry
  13. Hepatitis B Virus–Induced Imbalance of Inflammatory and Antiviral Signaling by Differential Phosphorylation of STAT1 in Human Monocytes
    Authors: Hongxiao Song, Guangyun Tan, Yang Yang, An Cui, Haijun Li, Tianyang Li et al.
    The Journal of Immunology
  14. Pro-Inflammatory Responses in Human Bronchial Epithelial Cells Induced by Spores and Hyphal Fragments of Common Damp Indoor Molds
    Authors: Elisabeth Øya, Rune Becher, Leni Ekeren, Anani K.J. Afanou, Johan Øvrevik, Jørn A. Holme
    International Journal of Environmental Research and Public Health
  15. Secretion of the Phosphorylated Form of S100A9 from Neutrophils Is Essential for the Proinflammatory Functions of Extracellular S100A8/A9
    Authors: Véronique Schenten, Sébastien Plançon, Nicolas Jung, Justine Hann, Jean-Luc Bueb, Sabrina Bréchard et al.
    Frontiers in Immunology
  16. Compound A attenuates toll-like receptor 4-mediated paclitaxel resistance in breast cancer and melanoma through suppression of IL-8
    Authors: R Sootichote, P Thuwajit, E Singsuksaw, M Warnnissor, PT Yenchitsom, S Ithimakin, J Chantharas, C Thuwajit
    BMC Cancer, 2018-02-27;18(1):231.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  17. Inositol-Requiring Enzyme 1-Mediated Downregulation of MicroRNA (miR)-146a and miR-155 in Primary Dermal Fibroblasts across ThreeTNFRSF1AMutations Results in Hyperresponsiveness to Lipopolysaccharide
    Authors: SR Harrison, T Scambler, L Oubussad, C Wong, M Wittmann, MF McDermott, S Savic
    Front Immunol, 2018-02-06;9(0):173.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  18. Tenascin C upregulates interleukin-6 expression in human cardiac myofibroblasts via toll-like receptor 4
    Authors: Azhar Maqbool, Emma J Spary, Iain W Manfield, Michaela Ruhmann, Lorena Zuliani-Alvarez, Filomena O Gamboa-Esteves et al.
    World Journal of Cardiology
  19. Induction of microglial toll-like receptor 4 by prothrombin kringle-2: a potential pathogenic mechanism in Parkinson’s disease
    Authors: Won-Ho Shin, Min-Tae Jeon, Eunju Leem, So-Yoon Won, Kyoung Hoon Jeong, Sang-Joon Park et al.
    Scientific Reports
  20. HIV-1 Tat Protein Induces Production of Proinflammatory Cytokines by Human Dendritic Cells and Monocytes/Macrophages through Engagement of TLR4-MD2-CD14 Complex and Activation of NF-kappaB Pathway.
    Authors: Ben Haij N, Planes R, Leghmari K, Serrero M, Delobel P, Izopet J, BenMohamed L, Bahraoui E
    PLoS ONE, 2015-06-19;10(6):e0129425.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  21. TLR4 Expression Is Associated with Left Ventricular Dysfunction in Patients Undergoing Coronary Artery Bypass Surgery.
    Authors: Avlas O, Bragg A, Fuks A, Nicholson J, Farkash A, Porat E, Aravot D, Levy-Drummer R, Cohen C, Shainberg A, Arad M, Hochhauser E
    PLoS ONE, 2015-06-01;10(6):e0120175.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  22. The C-terminal disulfide bonds of Helicobacter pylori GroES are critical for IL-8 secretion via the TLR4-dependent pathway in gastric epithelial cells.
    Authors: Su Y, Yang J, Lee H, Sheu F, Hsu C, Lin S, Chow L
    J Immunol, 2015-03-13;194(8):3997-4007.
    Species: Bacteria - Helicobacter pylori
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  23. Necrotic cell-derived high mobility group box 1 attracts antigen-presenting cells but inhibits hepatocyte growth factor-mediated tropism of mesenchymal stem cells for apoptotic cell death.
    Authors: Vogel S, Borger V, Peters C, Forster M, Liebfried P, Metzger K, Meisel R, Daubener W, Trapp T, Fischer J, Gawaz M, Sorg R
    Cell Death Differ, 2015-01-09;22(7):1219-30.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  24. Regulation of the innate immune response by fibronectin: synergism between the III-1 and EDA domains.
    Authors: Kelsh R, You R, Horzempa C, Zheng M, McKeown-Longo P
    PLoS ONE, 2014-07-22;9(7):e102974.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  25. Effect of thrombin on human amnion mesenchymal cells, mouse fetal membranes, and preterm birth.
    Authors: Mogami, Haruta, Keller, Patrick, Shi, Haolin, Word, R Ann
    J Biol Chem, 2014-03-20;289(19):13295-307.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  26. Pathogenic strains of Acanthamoeba are recognized by TLR4 and initiated inflammatory responses in the cornea.
    Authors: Alizadeh H, Tripathi T, Abdi M, Smith A
    PLoS ONE, 2014-03-14;9(3):e92375.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  27. Serum amyloid A in the placenta and its role in trophoblast invasion.
    Authors: Sandri S, Urban Borbely A, Fernandes I, de Oliveira E, Knebel F, Ruano R, Zugaib M, Filippin-Monteiro F, Bevilacqua E, Campa A
    PLoS ONE, 2014-03-10;9(3):e90881.
    Species: Human
    Sample Types: Whole Cells
    Applications: Functional Assay
  28. Activated platelets interfere with recruitment of mesenchymal stem cells to apoptotic cardiac cells via high mobility group box 1/Toll-like receptor 4-mediated down-regulation of hepatocyte growth factor receptor MET.
    Authors: Vogel S, Chatterjee M, Metzger K, Borst O, Geisler T, Seizer P, Muller I, Mack A, Schumann S, Buhring H, Lang F, Sorg R, Langer H, Gawaz M
    J Biol Chem, 2014-02-24;289(16):11068-82.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry, Neutralization
  29. Slit2-Robo4 pathway modulates lipopolysaccharide-induced endothelial inflammation and its expression is dysregulated during endotoxemia.
    Authors: Zhao H, Anand A, Ganju R
    J Immunol, 2013-11-22;192(1):385-93.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  30. Rattus Model Utilizing Selective Pulmonary Ischemia Induces Bronchiolitis Obliterans Organizing Pneumonia
    Authors: John C. Densmore, Paul M. Jeziorczak, Anne V. Clough, Kirkwood A. Pritchard, Breana Cummens, Meetha Medhora et al.
    Shock
  31. Fetal fibronectin signaling induces matrix metalloproteases and cyclooxygenase-2 (COX-2) in amnion cells and preterm birth in mice.
    Authors: Mogami H, Kishore A, Shi H, Keller P, Akgul Y, Word R
    J Biol Chem, 2012-11-26;288(3):1953-66.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  32. Toll-like receptor 4 mediates intrauterine growth restriction after systemic Campylobacter rectus infection in mice
    Authors: R.M. Arce, K.M. Caron, S.P. Barros, S. Offenbacher
    Molecular Oral Microbiology
  33. Protection by 20-5,14-HEDGE Against Surgically Induced Ischemia Reperfusion Lung Injury in Rats
    Authors: Irshad Ali, Stephanie Gruenloh, Ying Gao, Anne Clough, John R. Falck, Meetha Medhora et al.
    The Annals of Thoracic Surgery
  34. Isotretinoin increases skin-surface levels of neutrophil gelatinase-associated lipocalin in patients treated for severe acne.
    Authors: Lumsden KR, Nelson AM, Dispenza MC, Gilliland KL, Cong Z, Zaenglein AL, Thiboutot DM
    Br. J. Dermatol., 2011-08-01;165(2):302-10.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  35. 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.
    Species: Human
    Sample Types: Saliva
    Applications: Western Blot

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