Anti-TLR4 antibody [HTA125] - Low Endotoxin (ab30667)

Overview

  • Product nameAnti-TLR4 antibody [HTA125] - Low Endotoxin
    See all TLR4 primary antibodies
  • Description
    Mouse monoclonal [HTA125] to TLR4 - Low Endotoxin
  • Specificityab30667 recognises the human Toll like receptor 4 (TLR4) cell surface antigen. This antibody has been demonstrated to block activation of monocytes with LPS. TLR4 expression levels and cleavage or degradation products can vary between different cell and tissue samples. Customers have observed this variability in WB band size and our laboratory has confirmed this variability as well observing lower molecular weight cleavage and degradation products and in some samples a lack of the full length TLR4 band. The TLR4 cleavage and degradation products and potential lack of full length TLR4 are well documented in the literature, including PMID 16885150 and 22927440. We recommend running a positive control human intestine tissue lysate.
  • Tested applicationsSuitable for: WB, Flow Cyt, IP, Blocking, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat, Human, Rhesus monkey
  • Immunogen

    Ba/F3 cell line expressing TLR4.

  • General notesEndotoxin Level: <0.01 EU/ug

Properties

Applications

Our Abpromise guarantee covers the use of ab30667 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration. PubMed: 20622903
Flow Cyt 1/10 - 1/50.

Use 10µl of the suggested working dilution to label 106 cells or 100µl whole blood.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.
Blocking Use at an assay dependent concentration. PubMed: 17767165
ICC/IF Use at an assay dependent concentration. PubMed: 17916800

Target

  • FunctionCooperates with LY96 and CD14 to mediate the innate immune response to bacterial lipopolysaccharide (LPS). Acts via MYD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Also involved in LPS-independent inflammatory responses triggered by Ni(2+). These responses require non-conserved histidines and are, therefore, species-specific.
  • Tissue specificityHighly expressed in placenta, spleen and peripheral blood leukocytes. Detected in monocytes, macrophages, dendritic cells and several types of T-cells.
  • Involvement in diseaseGenetic variation in TLR4 is associated with age-related macular degeneration type 10 (ARMD10) [MIM:611488]. ARMD is a multifactorial eye disease and the most common cause of irreversible vision loss in the developed world. In most patients, the disease is manifest as ophthalmoscopically visible yellowish accumulations of protein and lipid that lie beneath the retinal pigment epithelium and within an elastin-containing structure known as Bruch membrane.
  • Sequence similaritiesBelongs to the Toll-like receptor family.
    Contains 18 LRR (leucine-rich) repeats.
    Contains 1 LRRCT domain.
    Contains 1 TIR domain.
  • DomainThe TIR domain mediates interaction with NOX4.
  • Post-translational
    modifications
    N-glycosylated. Glycosylation of Asn-526 and Asn-575 seems to be necessary for the expression of TLR4 on the cell surface and the LPS-response. Likewise, mutants lacking two or more of the other N-glycosylation sites were deficient in interaction with LPS.
  • Cellular localizationMembrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ARMD10 antibody
    • CD284 antibody
    • CD284 antigen antibody
    • Homolog of Drosophila toll antibody
    • hToll antibody
    • TLR 4 antibody
    • TLR4 antibody
    • TLR4_HUMAN antibody
    • TOLL antibody
    • Toll like receptor 4 antibody
    • Toll-like receptor 4 antibody
    see all

Anti-TLR4 antibody [HTA125] - Low Endotoxin images

  • ab30667 staining TLR4 in human peripheral blood monocytes by Flow Cytometry analysis.
  • ICC/IF image of ab30667 stained rat adrenal medulla PC12 cells. The cells were 4% PFA and 100% Methanol fixed (10 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab30667, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).

References for Anti-TLR4 antibody [HTA125] - Low Endotoxin (ab30667)

This product has been referenced in:
  • Garcia PV  et al. Increased toll-like receptors and p53 levels regulate apoptosis and angiogenesis in non-muscle invasive bladder cancer: mechanism of action of P-MAPA biological response modifier. BMC Cancer 16:422 (2016). WB . Read more (PubMed: 27389279) »
  • Wang XQ  et al. CLI-095 decreases atherosclerosis by modulating foam cell formation in apolipoprotein E-deficient mice. Mol Med Rep 14:49-56 (2016). Read more (PubMed: 27176130) »

See all 14 Publications for this product

Product Wall

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Immunogen is human TLR4-transfected mouse cell line Ba/F3, http://jem.rupress.org/content/189/11/1777.long

No epitope mapping has been performed, but based on application info for this antibody, it should be ...

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Thank you for contacting us.

The immunogen for ab30667, clone HTA125, was human TLR4-transfected mouse cell line Ba/F3, according to the paper describing development of the clone at the following URL, Shimazu et al, 1999:

http://j...

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Thank you for your inquiry.

I can confirm that the storage temperature of our products can be found on the individual datasheets.

Short term storage is usually two weeks. If the products is not used within two weeks of purchase, we ...

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Gracias por tu respuesta, y por compartir tus resultados con nosotros.

Es una lástima que este anticuerpo tampoco haya conseguido los resultados esperados.

Has probado ya dos lotes distintos de ab22048 y un lote de ab30677 si...

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Thank you for your question about ab30667. Generally, functional assays can cover a number of different experiments. However, this antibody has been used in blocking studies. As these will each be slightly different, we cannot provide a protocol, or gi...

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100 in PBS
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Dr. Kirk McManus

Verified customer

Submitted Feb 25 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"