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Anti-TLR3, clone PaT3, Cat. No. MABF2205, is a mouse monoclonal antibody that detects Toll-like receptor 3 and is tested for use in Flow Cytometry and Immunoprecipitation.
More>>Anti-TLR3, clone PaT3, Cat. No. MABF2205, is a mouse monoclonal antibody that detects Toll-like receptor 3 and is tested for use in Flow Cytometry and Immunoprecipitation. Less<<
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Übersicht
Replacement Information
Description
Catalogue Number
MABF2205-100UG
Description
Anti-TLR3 Antibody, clone PaT3
Alternate Names
Toll-like receptor 3
CD283
Background Information
Toll-like receptor 3 (UniProt: Q99MB1; also known as CD283, TLR3) is encoded by the Tlr3 gene (Gene ID: 142980) in murine species. Toll-like Receptors (TLRs) are transmembrane proteins that are expressed on various immune cells. They contain an N-terminal extracellular and a C-terminal cytoplasmic domain. The N-terminal region contains leucine-rich repeats (LRR) that recognize specific pathogen components. Although the ligand recognition mechanisms of the TLRs vary significantly, all TLR dimers exhibit similar overall arrangement. At least 13 different members of TLR family have been identified that detect different pathogen associated molecular patterns (PAMPs). TLR3 is a single-pass type I membrane glycoprotein that is highly expressed in lung tissue. It is synthesized with a signal peptide (aa 1-25), which is subsequently cleaved off to produce the mature form that contains a long extracellular domain (aa 26-705), a transmembrane domain (aa 706-726), and cytoplasmic domain (aa 727-925). It exists in monomeric or dimeric form and its dimerization is triggered by ligand binding. TLR3 is a nucleotide-sensing TLR that is activated by double-stranded RNA (dsRNA), a sign of viral infection. It contains 22 LRR repeats and its ds-RNA binding is mediated by LRR 1-3 and 17,18. TLR3 acts via the adapter TRIF/TICAM1, leading to NF-kappa-B activation, IRF3 nuclear translocation, cytokine secretion and the inflammatory response. Clone Pat3 is specific for murine TLR3 and does not react with TLR7, 8, or 9. However, it reacts with both the cleaved and uncleaved forms of TLR3. (Ref.: Murakami, Y., et al. (2014). J. Immunol. 193 (10); 5208-5217).
References
Product Information
Format
Purified
Presentation
Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Anti-TLR3, clone PaT3, Cat. No. MABF2205, is a mouse monoclonal antibody that detects Toll-like receptor 3 and is tested for use in Flow Cytometry and Immunoprecipitation.
Key Applications
Flow Cytometry
Immunoprecipitation
Application Notes
Immunoprecipitation Analysis: A representative lot immunoprecipitated TLR3 in Immunoprecipitation applications (Murakami, Y., et. al. (2014). J Immunol. 193(10):5208-17).
Flow Cytometry Analysis: A representative lot detected TLR3 in Flow Cytometry applications (Pelka, K., et. al. (2018). Immunity. 48(5):911-922; Murakami, Y., et. al. (2014). J Immunol. 193(10):5208-17).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Biological Information
Immunogen
Ba/F3 cells expressing Flag-6×His-conjugated mouse TLR3. Tlr3-/- mice were immunized.
Epitope
Unknown
Clone
PaT3
Concentration
1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Host
Mouse
Specificity
Clone PaT3 is a mouse monoclonal antibody that specifically detects murine Toll-like receptor 3.
Flow Cytometry Analysis: 2 µg of this antibody detected TLR3 in one million J774.2 cells.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at +2°C to +8°C from date of receipt.
Unc-93 homolog B1 (UNC93B1) is a key regulator of nucleic acid (NA)-sensing Toll-like receptors (TLRs). Loss of NA-sensing TLR responses in UNC93B1-deficient patients facilitates Herpes simplex virus type 1 (HSV-1) encephalitis. UNC93B1 is thought to guide NA-sensing TLRs from the endoplasmic reticulum (ER) to their respective endosomal signaling compartments and to guide the flagellin receptor TLR5 to the cell surface, raising the question of how UNC93B1 mediates differential TLR trafficking. Here, we report that UNC93B1 regulates a step upstream of the differential TLR trafficking process. We discovered that UNC93B1 deficiency resulted in near-complete loss of TLR3 and TLR7 proteins in primary splenic mouse dendritic cells and macrophages, showing that UNC93B1 is critical for maintaining TLR expression. Notably, expression of an ER-retained UNC93B1 version was sufficient to stabilize TLRs and largely restore endosomal TLR trafficking and activity. These data are critical for an understanding of how UNC93B1 can regulate the function of a broad subset of TLRs.
TLR3 senses viral dsRNA in endolysosomes. The TLR3 ectodomain is cleaved by proteases such as cathepsins in endolysosomes. It remains controversial whether the N-terminal fragment of TLR3 ectodomain (TLR3N) is cleaved off or remains associated with the C-terminal TLR3 fragment (TLR3C). In addition to endosomes, TLR3 is reported to be expressed on the surface of human fibroblasts, but not of human monocyte-derived dendritic cells. Less is known about roles of TLR3N and cell surface TLR3 in dsRNA sensing. In this study, we show the cleavage site of the TLR3 ectodomain and cell surface expression of TLR3 on mouse primary immune cells. TLR3C, which started at 343S, was associated with TLR3N. Both TLR3N and TLR3C were required for activation of IFN-β and NF-κB promoters by dsRNA, demonstrating that dsRNA is sensed by the TLR3N+C complex. Newly established mAbs to mouse TLR3 revealed that cell surface TLR3 was highly expressed on splenic CD8(+) dendritic cells and marginal zone B cells. Cell surface expression of TLR3 on these cells was dependent on the TLR-specific transporter Unc93B1. Although cell surface TLR3 was only weakly expressed on macrophages, TLR3 mAb specifically enhanced TLR3 responses to dsRNA. These results demonstrate that dsRNA is sensed by the TLR3N+C complex and that cell surface TLR3 is a promising target for modulating TLR3 responses.