TLR4 Antibody (HTA125) [FITC]

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Flow Cytometry: TLR4 Antibody (HTA125) [FITC] [NB100-56059] - An intracellular stain was performed on MG-63 cells with TLR4 Antibody [HTA125] NB100-56059 (blue) and a matched isotype control (orange). Cells were fixed ...read more

Product Details

Summary
Reactivity Hu, Mu, CaSpecies Glossary
Applications Flow, Flow-CS, Flow-IC, ICC/IF, IHC, WB (-)
Clone
HTA125
Clonality
Monoclonal
Host
Mouse
Conjugate
FITC

Order Details

TLR4 Antibody (HTA125) [FITC] Summary

Immunogen
This TLR4 Antibody (HTA125) [FITC] was developed by immunizing mice with Ba/F3 cell line expressing human TLR4 cell surface antigen.
Specificity
The antibody has been shown to block the activation of monocytes with LPS (Paik, et al, 2003).
Isotype
IgG2a
Clonality
Monoclonal
Host
Mouse
Gene
TLR4
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Flow Cytometry 1uL / 1 million cells
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry
  • Flow (Cell Surface)
  • Flow (Intracellular)
Application Notes
Confocal Microscopy: see Scheel et al. (2006) for details. FA (Neutralization): see Basek et al, 2005 for details. Flow (Cell Surface): 2-5 ug / 1x10^6 cells, IF/ICC: please see Schneeman et al (2005) for details. IP: please see Shimazu et al, 1999 for details. Use in Immunohistochemistry and Immunocytochemistry/immunofluorescence reported in scientific literature (PMID: 28219705). Use in Flow-intracellular reported in scientific literature (PMID: 18295348). Use in Flow-cell surface reported in scientific literature (PMID: 17015691).
Theoretical MW
95.7 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Agonist
Antagonist
Publications
Read Publications using
NB100-56059 in the following applications:

Packaging, Storage & Formulations

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

Alternate Names for TLR4 Antibody (HTA125) [FITC]

  • 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.

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Publications for TLR4 Antibody (NB100-56059)(26)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 6 applications: FLOW, Flow, Flow-CS, Flow-Cs, Flow-IC, ICC/IF.


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(3)
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(1)
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Showing Publications 1 - 10 of 26. Show All 26 Publications.
Publications using NB100-56059 Applications Species
Akhter N, Madhoun A, Arefanian H et al. Oxidative Stress Induces Expression of the Toll-Like Receptors (TLRs) 2 and 4 in the Human Peripheral Blood Mononuclear Cells: Implications for Metabolic Inflammation Cell. Physiol. Biochem. Jun 3 2019 [PMID: 31162913] (Flow, Human) Flow Human
Ahmad Rasheed, Al-Roub Areej, Kochumon Shihab et al. The Synergy between Palmitate and TNF-a for CCL2 Production Is Dependent on the TRIF/IRF3 Pathway: Implications for Metabolic Inflammation. Journal of Immunology (Baltimore, Md. : 1950) 2018 [PMID: 29632147] (Flow, Human) Flow Human
Heftrig D, Sturm R, Oppermann E et al. Impaired Surface Expression of HLA-DR, TLR2, TLR4, and TLR9 in Ex Vivo-In Vitro Stimulated Monocytes from Severely Injured Trauma Patients.. Mediators Inflamm. Mar 3 2017 [PMID: 28255201]
Larsson O, Tengroth L, Xu Y et al. Substance P represents a novel first-line defense mechanism in the nose. J. Allergy Clin. Immunol. Feb 20 2017 [PMID: 28219705] (Flow, Mouse) Flow Mouse
Jamin A, Dehoux L, Dossier C et al. Toll-like receptor 3 expression and function in childhood idiopathic nephrotic syndrome Clin. Exp. Immunol. 2015 Jun 30 [PMID: 26123900] (FLOW, Human)

Details:
TLR4 antibody clone HTA125 [FITC] (Imgenex: IMG-417C) was used for FLOW on peripheral blood mononuclear cells/PBMC of idiopathic nephrotic syndrome/INS children, including 28 in relapse, 23 in remission and 40 controls. The cells were incubated with conjugated antibodies along with isotype control for 30 minutes at 4C and 10(5) viable PBMCs were analyzed using FACSCantoll/FACSDiva.
FLOW Human
Harman AN, Bye CR, Nasr N et al. Identification of lineage relationships and novel markers of blood and skin human dendritic cells. J Immunol. 2013 Jan 1 [PMID: 23183897]
Matsunaga N, Tsuchimori N, Matsumoto T, Ii M. TAK-242 (resatorvid), a small-molecule inhibitor of Toll-like receptor (TLR) 4 signaling, binds selectively to TLR4 and interferes with interactions between TLR4 and its adaptor molecules. Mol Pharmacol. 2011 Jan [PMID: 20881006]
Pillay J, Ramakers BP, Kamp VM et al. Functional heterogeneity and differential priming of circulating neutrophils in human experimental endotoxemia. J Leukoc Biol. 2010 Jul [PMID: 20400675]
Wu CY, Chi PL, Hsieh HL et al. TLR4-dependent induction of vascular adhesion molecule-1 in rheumatoid arthritis synovial fibroblasts: Roles of cytosolic phospholipase A(2)alpha/cyclooxygenase-2. J Cell Physiol. 2010 May [PMID: 20112284]
Pietschmann K, Beetz S, Welte S et al. Toll-like receptor expression and function in subsets of human gammadelta T lymphocytes. Scand J Immunol. 2009 Sep [PMID: 19703014]
Show All 26 Publications.

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Product General Protocols

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Video Protocols

ICC/IF Video Protocol

FAQs for TLR4 Antibody (NB100-56059). (Showing 1 - 2 of 2 FAQs).

  1. I would like to use this antibody but it has not been validated in my species of interest. Is there any way I can find out if it will work?
    • We offer risk-free testing of all of our primary antibodies. Please check out our Innovator's Reward Program and test this TLR4 antibody in any unvalidated species or application, without the financial risk of failure.
  2. How do I choose secondary antibodies to label the same cells when I have two primary antibodies from the same host?
    • Use isotype-specific secondary antibodies if the primary antibodies are of different isotypes. You can also make direct conjugates of the primary antibodies by use of antibody labeling kits, dyes, or custom conjugations (please contact Technical Support for custom orders).

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NB100-56723AF350
Alexa Fluor 405 NB100-56727AF405
Alexa Fluor 405 NB100-56723AF405
Alexa Fluor 488 NBP2-24897
Alexa Fluor 532 NB100-56723AF532
Alexa Fluor 594 NB100-56723AF594
Alexa Fluor 647 NB100-56727AF647
Alexa Fluor 647 NB100-56723AF647
Alexa Fluor 700 NB100-56727AF700
Alexa Fluor 700 NB100-56723AF700
Alexa Fluor 750 NB100-56723AF750
Allophycocyanin NB100-56727APC
Allophycocyanin NB100-56723APC
Allophycocyanin/Cy7 NB100-56723APCCY7
Biotin NBP2-24744
DyLight 350 NB100-56723UV
DyLight 350 NB100-56727UV
DyLight 405 NB100-56723V
DyLight 405 NB100-56727V
DyLight 488 NB100-56727G
DyLight 488 NB100-56723G
DyLight 550 NB100-56727R
DyLight 594 NB100-56723DL594
DyLight 650 NB100-56727C
DyLight 680 NB100-56727FR
DyLight 680 NB100-56723FR
DyLight 755 NB100-56723IR
DyLight 755 NB100-56727IR
FITC NB100-56059
Janelia Fluor 549 NB100-56723JF549
Janelia Fluor 549 NB100-56727JF549
Janelia Fluor 646 NB100-56727JF646
Janelia Fluor 646 NB100-56723JF646
PE NB100-56062
PE/Atto594 NB100-56723PEATT594
PE/Cy5.5 NB100-56723PECY55
PE/Cy7 NB100-56723PECY7
PerCP NB100-56723PCP
PerCP NB100-56727PCP

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Blogs on TLR4. Showing 1-10 of 11 blog posts - Show all blog posts.

PAMPs and DAMPs: What is the same and what is different about these molecules?
By Victoria OsinskiWhat are PAMPs and DAMPsInflammation results from stimuli signaling damage or infection. The inflammatory response can be beneficial or harmful depending on the type and duration of stimuli. The s...  Read full blog post.

How to identify B cell subsets using flow cytometry
By Victoria OsinskiUsing flow cytometry to identify B cell subsetsIdentifying cellular subsets by flow cytometry requires careful and thorough planning in order to ensure the correct subset of cells are identified a...  Read full blog post.

Lipopolysaccharide from gut microbiome localizes in human atherosclerotic plaques and promotes TLR4-mediated oxidative stress
By Jamshed Arslan, Pharm. D., PhD. Atherosclerosis is a chronic inflammatory condition in which plaques of fats and other substances slowly buildup on the inner walls of arteries to restrict blood flow. In atheroscle...  Read full blog post.

Toll-like receptors in the intestinal epithelial cells
By Jamshed Arslan, Pharm. D., PhD. Toll-like receptors (TLRs) are microbe-sensing proteins that act as first responders to danger signals. TLRs help the intestinal epithelial cells (IECs) recognize commensal bacteria...  Read full blog post.

The role of STING/TMEM173 in gamma and encephalitis Herpes Simplex Virus (HSV)
Stimulator of interferon genes (STING), also known as TMEM173, promotes the production of the interferon’s IFN-alpha and IFN-beta.  STING possesses three functional domains: a cytoplasmic C-terminal tail, a central globular domain, and four N-...  Read full blog post.

TRIF/TICAM1 and mitochondrial dynamics in the innate immune response
TRIF, also known as toll like receptor adaptor molecule 1 or TICAM1, is known for its role in invading foreign pathogens as part of our innate immune response. TRIF/TICAM1 is a TIR-domain adaptor protein (toll/interleukin-1 receptor) that interacts...  Read full blog post.

The role of TLR4 in breast cancer
Toll like receptors (TLRs) are highly conserved proteins that are first known for their role in pathogen recognition and immune response activation.  In order to elicit the necessary immune response in reaction to a foreign pathogen, TLRs trigger cy...  Read full blog post.

cIAP2 - balancing cell death and cell survival
The inhibitor of apoptosis proteins (IAPs) are important regulators of cell death and inflammation. The cellular inhibitor of apoptosis protein 2 (cIAP2) contains three Baculovirus IAP repeat (BIR) domains, a Ubiquitin associated (UBA) domain, and...  Read full blog post.

TLR4 - A Guardian of Innate Immunity
Toll-like receptor 4 (TLR4) belongs to the family of Toll-like receptors (TLR), and plays a main role in pathogen recognition and innate immunity system activation. The TLR family members are highly conserved proteins that all contain a high degree of...  Read full blog post.

IRAK4: The "master IRAK" critical for initiating immune responses
IRAK4, also known as Interleukin-1 receptor-associated kinase 4, is a serine/threonine-protein kinase that plays a critical role in initiating innate and adaptive immune responses against foreign pathogens. It activates NF-kappaB in both Toll-like rec...  Read full blog post.

Showing 1-10 of 11 blog posts - Show all blog posts.
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Bioinformatics

Gene Symbol TLR4