TLR3 Antibody - BSA Free

Novus Biologicals | Catalog # NB100-56571

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown, Orthogonal Validation, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Porcine

Cited:

Human, Mouse

Predicted:

Rat (91%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Flow (Cell Surface), Flow (Intracellular), Dot Blot, Knockdown Validated

Cited:

Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Flow (Cell Surface), Cytometric Bead Assay Standard, Flow Cytometry Control

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
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Product Specifications

Immunogen

A mix of synthetic peptides corresponding to amino acids 135-150 (SIHKIKSNPFKNQKNL), 828-844 (CRRFKVHHAVQQAIEQN), and 876-891 (CILNWPVQKERINAFH) of mouse TLR3.

Reactivity Notes

Predicted to cross-react with Rat. Porcine data from customer review.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

104 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.

Description

Novus Biologicals Rabbit TLR3 Antibody - BSA Free (NB100-56571) is a polyclonal antibody validated for use in IHC, WB and Flow. Anti-TLR3 Antibody: Cited in 18 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for TLR3 Antibody - BSA Free

Western Blot: TLR3 Antibody [NB100-56571]

Western Blot: TLR3 Antibody [NB100-56571]

TLR3-Antibody-Western-Blot-NB100-56571-img0009.jpg
Immunohistochemistry-Frozen: TLR3 Antibody [NB100-56571]

Immunohistochemistry-Frozen: TLR3 Antibody [NB100-56571]

Immunohistochemistry-Frozen: TLR3 Antibody [NB100-56571] - Frozen mouse eye tissue sections stained with TLR3, a retinal pigment epithelium (RPE) marker and DAPI. Top panel: TLR3 (red) colocalized with the RPE (green) marker. Bottom panel: Tissue stained with isotype control (no TLR3 Ab), RPE and DAPI. Data courtesy of Kleinman et al.
Flow Cytometry: TLR3 Antibody [NB100-56571]

Flow Cytometry: TLR3 Antibody [NB100-56571]

Flow Cytometry: TLR3 Antibody [NB100-56571] - Analysis using the FITC conjugate of NB100-56571. Staining of TLR3 in Balb/c mouse splenocytes using this antibody at 1 ug/10^6 cells. Green represents rabbit IgG isotype control this antibody; red represents anti-TLR3 antibody.
Western Blot: TLR3 Antibody [NB100-56571]

Western Blot: TLR3 Antibody [NB100-56571]

Western Blot: TLR3 Antibody [NB100-56571] - Analysis of TLR3 in mouse spleen tissue lysate using NB100-56571 at 2 ug/ml.
Immunohistochemistry: TLR3 Antibody [NB100-56571]

Immunohistochemistry: TLR3 Antibody [NB100-56571]

Immunohistochemistry: TLR3 Antibody [NB100-56571] - Pig lung poisoned through aspiration with gastric juice and organophosphate (dimethoate EC40) after 48 hours of IC ventilation. Image from verified customer review.
Flow Cytometry: TLR3 Antibody [NB100-56571]

Flow Cytometry: TLR3 Antibody [NB100-56571]

Flow Cytometry: TLR3 Antibody [NB100-56571] - Intracellular flow analysis of TLR3 in Balb/c mouse splenocytes and mouse RAW cells using TLR3 antibody (red) and isotype control antibody (green) at 1 ug/10^6 cells.
Knockdown Validated: TLR3 Antibody [NB100-56571]

Western Blot: TLR3 Antibody [NB100-56571]

TLR3-Antibody-Knockdown-Validated-NB100-56571-img0010.jpg
TLR3 Antibody

Western Blot: TLR3 Antibody [NB100-56571] -

Western Blot: TLR3 Antibody [NB100-56571] - Myocardial infarction (MI) increased TLR3 expression & its physical association with Trif in mouse myocardium. Heart samples were taken from infarct area at 4 weeks after MI. (A) & (B) show mRNA & protein levels of TLR3 in sham & MI hearts. n = 4 mice/group. Data are means ± S.D. AP < 0.01 versus sham. (C) Representative immunohistochemistry images of heart sections stained for TLR3 (brown colour). An isotype IgG control was performed to verify the specificity of TLR3 reactivity. (D) Lysates of heart tissue were immunoprecipitated with anti‐TLR3 antibodies (IP: TLR3), followed by SDS–PAGE & immunoblotting (IB) with indicated antibodies. IP with isotype IgG (IP: IgG) was performed as a control to exclude the non‐specific binding of antibodies to cellular proteins. Green arrows indicate non‐specific bands. The association between TLR3 & Trif, but not MyD88, was detectable in sham myocardium & was increased in infarct myocardium. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28945004), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TLR3 Antibody

Western Blot: TLR3 Antibody [NB100-56571] -

Western Blot: TLR3 Antibody [NB100-56571] - Myocardial infarction (MI) increased TLR3 expression & its physical association with Trif in mouse myocardium. Heart samples were taken from infarct area at 4 weeks after MI. (A) & (B) show mRNA & protein levels of TLR3 in sham & MI hearts. n = 4 mice/group. Data are means ± S.D. AP < 0.01 versus sham. (C) Representative immunohistochemistry images of heart sections stained for TLR3 (brown colour). An isotype IgG control was performed to verify the specificity of TLR3 reactivity. (D) Lysates of heart tissue were immunoprecipitated with anti‐TLR3 antibodies (IP: TLR3), followed by SDS–PAGE & immunoblotting (IB) with indicated antibodies. IP with isotype IgG (IP: IgG) was performed as a control to exclude the non‐specific binding of antibodies to cellular proteins. Green arrows indicate non‐specific bands. The association between TLR3 & Trif, but not MyD88, was detectable in sham myocardium & was increased in infarct myocardium. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28945004), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TLR3 Antibody

Immunohistochemistry: TLR3 Antibody [NB100-56571] -

Immunohistochemistry: TLR3 Antibody [NB100-56571] - Myocardial infarction (MI) increased TLR3 expression & its physical association with Trif in mouse myocardium. Heart samples were taken from infarct area at 4 weeks after MI. (A) & (B) show mRNA & protein levels of TLR3 in sham & MI hearts. n = 4 mice/group. Data are means ± S.D. AP < 0.01 versus sham. (C) Representative immunohistochemistry images of heart sections stained for TLR3 (brown colour). An isotype IgG control was performed to verify the specificity of TLR3 reactivity. (D) Lysates of heart tissue were immunoprecipitated with anti‐TLR3 antibodies (IP: TLR3), followed by SDS–PAGE & immunoblotting (IB) with indicated antibodies. IP with isotype IgG (IP: IgG) was performed as a control to exclude the non‐specific binding of antibodies to cellular proteins. Green arrows indicate non‐specific bands. The association between TLR3 & Trif, but not MyD88, was detectable in sham myocardium & was increased in infarct myocardium. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28945004), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TLR3 Antibody

Western Blot: TLR3 Antibody [NB100-56571] -

Western Blot: TLR3 Antibody [NB100-56571] - TLR3‐knockout (TLR3‐KO) attenuated cardiac autophagy induced by MI. The infarct & remote tissues were separately sampled from the left ventricle after 4 weeks of MI. Accordingly, anterior & posterior tissues of the left ventricle were sampled from sham hearts as controls. (A) Representative Western blot images & quantitative analyses of autophagy markers. n = 4–8/group. Quantitative data are fold changes of WT‐sham. aP < 0.05, AP < 0.01 versus respective WT‐sham; bP < 0.05, BP < 0.01 versus respective WT‐MI. (B) Representative electron microphotographs of ultrathin sections of resin‐embedded heart biopsies. Arrows indicate autophagic vacuoles. (C) Lysates of infarct tissue were immunoprecipitated (IP), followed by SDS–PAGE & immunoblotting (IB) with indicated antibodies. IP with isotype IgG served as a control. Green arrows indicate non‐specific bands. Representative images were taken from four independent experiments. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28945004), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TLR3 Antibody

Western Blot: TLR3 Antibody [NB100-56571] -

Western Blot: TLR3 Antibody [NB100-56571] - A TLR3 agonist polyinosinic‐polycytidylic acid (poly(I:C)) induced autophagy in cultured cardiomyocytes through a TRIF‐dependent pathway. (A) Poly(I:C) increased autophagy markers in cultured H9c2 rat ventricular cells. (B) Poly(I:C) stimulated autophagosome formation but did not affect autophagic flux. Primary cultured neonatal rat ventricular myocytes (NRVMs) were transfected with a tandem mRFP‐GFP‐LC3 adenovirus for 24 hrs, followed by treatment with poly(I:C) (100 μg/ml, 4 hrs). Autophagosomes & autolysosomes were, respectively, visualized as yellow‐ & red‐only punctas under a confocal microscope. (C) An autophagic flux inhibitor chloroquine (CQ) induced accumulations of LC3‐II & p62/SQSTM1 proteins in H9c2 myocytes receiving poly(I:C) (100 μg/ml, 4 hrs). CQ was applied at 10 μM, immediately prior to poly(I:C). (D) Effects of indicated siRNA on poly(I:C)‐induced changes in autophagy markers in NRVMs. All the siRNAs were transfected at 50 nM for 48 hrs, & poly(I:C) was added at 100 μg/ml for 4 hrs before cell harvest. Negative control (NC) siRNA served as control. RNAiMAX transfection reagent was used in all the siRNA experiments. The upper panel shows the knockdown effects of siRNAs, & the lower panel shows representative Western blot images (presented from four independent experiments) & densitometry quantitative data (normalized into ‘fold of vehicle group’). All quantitative data are expressed as means ± S.D. aP < 0.05, AP < 0.01 versus vehicle; bP < 0.05, BP < 0.01 versus poly(I:C). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28945004), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for TLR3 Antibody - BSA Free

Application
Recommended Usage

Dot Blot

reported in scientific literature (PMID 27248820)

Flow (Cell Surface)

reported in scientific literature (Pawar et al (2006))

Flow (Intracellular)

1 ug/1 million cells

Flow Cytometry

1 ug/10^6 cells

Immunohistochemistry

1:20-1:1000

Immunohistochemistry-Frozen

1:20-1:1000. Use reported in scientific literature (Pawar et al (2005))

Immunohistochemistry-Paraffin

1:100. Use reported in scientific literature (Patole et al (2005))

Western Blot

1-3 ug/ml
Application Notes
Use in Flow (Intracellular) reported in multiple pieces of scientific literature.

Reviewed Applications

Read 1 review rated 4 using NB100-56571 in the following applications:

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: TLR3

Toll-like receptor 3 (TLR3) is a type I transmembrane glycoprotein that contributes to the innate immune response, recognizing distinct pathogen-associate molecular patterns (PAMPs) and damage-associate molecular patterns (DAMPs) (1,2). The TLR family member TLR3 specifically recognizes and binds double-stranded RNA (dsRNA) from viruses and the synthetic analog polyriboinosinic:polyribocytidylic acid (poly(I:C)) (1-5). TLR3 is typically expressed in the endosomes of innate immune cells including macrophages, natural killer (NK) cells, and dendritic cells (DCs) (1-5). TLR3 is also localized on the cell surface of fibroblasts, epithelial cells, and vascular endothelial cells (1-5). The human TLR3 protein is 904 amino acids (aa) in length with a theoretical molecular weight (MW) of 104 kDa (6). It consists of a 23 aa signal sequence, a horseshoe-shaped 681 aa extracellular domain (ECD) containing 23 leucine-rich repeats (LRRs), a 21 aa helical transmembrane domain, and a 179 aa cytoplasmic region containing a Toll/IL-1 receptor (TIR) domain (1,6). Upon ligand binding, TLR3-ECD dimerizes and the adapter protein TIR-domain-containing adapter inducing interferon-beta (TRIF/TICAM1) is recruited (1-5). TRIF interacts with tumor necrosis factor receptor-associated factor 3 (TRAF3) and TRAF6 and results in a signal transduction cascade involving activation of transcription factors interferon regulatory factor 3 (IRF3), IRF7, nuclear factor-kappaB (NF-kappaB), and activation protein-1 (AP-1) (1-5). Transcription factors translocate to the nucleus, driving type I interferon (IFN) production, secretion of pro-inflammatory cytokines, and tumor regression (1-5). Furthermore, TRIF can also interact with receptor-interacting serine-threonine kinase 1 (RIP1) and RIP3 leading to reactive oxygen species (ROS) production and apoptosis (1,2,5). Conversely, NF-kappaB transcription can also promote chemokine production and promote the WNT pathway associated with stemness and pro-tumorigenic properties (1,5).

Given the role of TLR3 in immune response, its expression or dysfunction has been associated with a number of pathologies from chronic inflammation to autoimmune disorders and cancer (1-5,7). TLR3 is expressed in many cancer types, often related to viral infection, such as cervical cancer, hepatocellular carcinoma (HCC), melanoma, breast cancer, and prostate cancer (1,5). TLR3 signaling has a dual role in cancer, either contributing to pro- or anti-tumor properties depending on the type of cancer (1,5). Therapeutic targeting the TLR3 signaling pathway is under investigation. TLR3 inhibitors or antagonists are being studied for the treatment autoimmune and inflammatory disorders such as of sepsis and atherosclerosis (2,8). TLR3 agonists, either alone or in combination with immune checkpoint inhibitors or therapeutic agents, are being studied as immunotherapeutic treatments of many cancers such as colorectal cancer, prostate cancer, and melanoma (7).

References

1. Zheng X, Li S, Yang H. Roles of Toll-Like Receptor 3 in Human Tumors. Front Immunol. 2021;12:667454. https://doi.org/10.3389/fimmu.2021.667454

2. Zhuang C, Chen R, Zheng Z, Lu J, Hong C. Toll-Like Receptor 3 in Cardiovascular Diseases. Heart Lung Circ. 2022;S1443-9506(22)00080-4. https://doi.org/10.1016/j.hlc.2022.02.012

3. Bianchi F, Pretto S, Tagliabue E, Balsari A, Sfondrini L. Exploiting poly(I:C) to induce cancer cell apoptosis. Cancer Biol Ther. 2017;18(10):747-756. https://doi.org/10.1080/15384047.2017.1373220

4. Matsumoto M, Seya T. TLR3: interferon induction by double-stranded RNA including poly(I:C). Adv Drug Deliv Rev. 2008;60(7):805-812. https://doi.org/10.1016/j.addr.2007.11.005

5. Muresan XM, Bouchal J, Culig Z, Soucek K. Toll-Like Receptor 3 in Solid Cancer and Therapy Resistance. Cancers (Basel). 2020;12(11):3227. https://doi.org/10.3390/cancers12113227

6. Uniprot (O15455)

7. Le Naour J, Galluzzi L, Zitvogel L, Kroemer G, Vacchelli E. Trial watch: TLR3 agonists in cancer therapy. Oncoimmunology. 2020;9(1):1771143. https://doi.org/10.1080/2162402X.2020.1771143

8. Gao W, Xiong Y, Li Q, Yang H. Inhibition of Toll-Like Receptor Signaling as a Promising Therapy for Inflammatory Diseases: A Journey from Molecular to Nano Therapeutics. Front Physiol. 2017;8:508. https://doi.org/10.3389/fphys.2017.00508

Long Name

Toll-like Receptor 3

Alternate Names

CD283

Entrez Gene IDs

7098 (Human); 142980 (Mouse)

Gene Symbol

TLR3

UniProt

Additional TLR3 Products

Product Documents for TLR3 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for TLR3 Antibody - BSA Free

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.

Citations for TLR3 Antibody - BSA Free

Customer Reviews for TLR3 Antibody - BSA Free (1)

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  • Name: Bryan Tinsley
    Application: Immunohistochemistry
    Sample Tested:
    Species: Other
    Verified Customer | Posted 09/18/2014
    pig lung poisoned through aspiration with gastric juice and organophosphate (dimethoate EC40) after 48 hours of IC ventilation.
    TLR3 Antibody - BSA Free NB100-56571

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