TLR3 Antibody - BSA Free

Western Blot: TLR3 Antibody [NB100-56571] - Myocardial infarction (MI) increased TLR3 expression and its physical association with Trif in mouse myocardium. Heart samples were taken from infarct area at 4 weeks after MI. (A) and (B) show mRNA and protein levels of TLR3 in sham and MI hearts. n = 4 mice/group. Data are means +/- S.D. AP < 0.01 versus sham. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1111/jcmm.13328) licensed under a CC-BY license.

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] - 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] - Analysis of TLR3 in mouse spleen tissue lysate using NB100-56571 at 2 ug/ml.

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] - 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.

Western Blot: TLR3 Antibody [NB100-56571] - siRNA was transfected at 50 nM for 48 hrs, and poly(I:C) was added at 100 ug/ml for 4 hrs before cell harvest. Negative control (NC) siRNA served as control. RNAiMAX transfection reagent was used in all the siRNA experiments. Panel shows the knockdown effects of siRNAs. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1111/jcmm.13328) licensed under a CC-BY license.

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 (//pubmed.ncbi.nlm.nih.gov/28945004), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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 (//pubmed.ncbi.nlm.nih.gov/28945004), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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 (//pubmed.ncbi.nlm.nih.gov/28945004), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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 (//pubmed.ncbi.nlm.nih.gov/28945004), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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 (//pubmed.ncbi.nlm.nih.gov/28945004), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Po, Rt
• Applications: WB, DB, Flow, IHC, KD
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

TLR3 Antibody - BSA Free Summary

• 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.
• Immunogen: A mix of synthetic peptides corresponding to amino acids 135-150 (SIHKIKSNPFKNQKNL), 828-844 (CRRFKVHHAVQQAIEQN), and 876-891 (CILNWPVQKERINAFH) of mouse TLR3.
• Predicted Species: Rat (91%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TLR3
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • 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))
  • Knockdown Validated
  • Western Blot 1-3 ug/ml
• Application Notes: Use in Flow (Intracellular) reported in multiple pieces of scientific literature.
• 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.
• Agonist:
  • Polyinosinic-polycytidylic acid HMW NBP2-25288
• Control:

  Mouse Spleen Whole Tissue Lysate (Adult Whole Normal)

Reactivity Notes

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

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for TLR3 Antibody - BSA Free

• CD283 antigen
• CD283
• TLR3
• toll-like receptor 3

Background

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

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.

Publications for TLR3 Antibody (NB100-56571)(19)

Publications using NB100-56571

• Cipollini OQ Investigating Microglial Innate Immune Responses to Amyloid-Beta (1-42) and Murine Betacoronavirus (MHV-A59) Thesis 1905-07-13 (WB)
• Min KW, Choi KM, Mun H et Al. Mature microRNA-binding protein QKI suppresses extracellular microRNA let-7b release J Cell Sci 2024-11-01 [PMID: 39308343]
• Do?an G, Sand?kç? M, Karagenç L. et Al. Stage-specific expression of Toll-like receptors in the seminiferous epithelium of mouse testis Histochem Cell Biol 2024-07-31 [PMID: 39085445]
• Nie H, Mei Z et al. Bushen recipe and its disassembled prescriptions inhibit inflammation of liver injury associated with Concanavalin A through Toll like receptor 3/9 signaling pathway. Mol Med Rep 2018-01-08 [PMID: 29845244] (WB, Mouse)
• Gao T, Zhang SP, Wang JF et al. TLR3 contributes to persistent autophagy and heart failure in mice after myocardial infarction J. Cell. Mol. Med. 2017-09-25 [PMID: 28945004] (WB, Mouse)
• Jeon YK, Kim CK, Koh J et al. Pellino-1 confers chemoresistance in lung cancer cells by upregulating cIAP2 through Lys63-mediated polyubiquitination. Oncotarget. 2016-07-05 [PMID: 27248820] (Cytometric Bead Assay Standard, Human)
• Jiang M, Broering R, Trippler M et al. MicroRNA-155 controls Toll-like receptor 3- and hepatitis C virus-induced immune responses in the liver. J. Viral Hepat. 2014-02-01 [PMID: 24383923] (Flow-CS, Mouse)
  
  Details:
  BalbC lymphocytes, Fig S1C. Expression of TLR3 was detected using unconjugated anti-TLR3 antibodies and secondary antibodies.
• Priya R, Dhanwani R, Patro IK et al. Differential regulation of TLR mediated innate immune response of mouse neuronal cells following infection with novel ECSA genotype of Chikungunya virus with and without E1:A226V mutation. Infect Genet Evol 2013-12-01 [PMID: 24126361] (WB, Mouse)
• Yamashita M, Chattopadhyay S, Fensterl V et al. Epidermal growth factor receptor is essential for Toll-like receptor 3 signaling. Sci Signal. 2012-07-17 [PMID: 22810896]
• Chen G Y, Hwang S M et al. Defective antiviral responses of induced pluripotent stem cells to baculoviral vector transduction. J Virol 2012-01-08 [PMID: 22623765] (FLOW, Mouse)
• Kleinman ME, Kaneko H, Cho WG et al. Short-interfering RNAs induce retinal degeneration via TLR3 and IRF3. Mol Ther. 2012-01-01 [PMID: 21988875]
  
  Details:
  Antibodies cited: 1. TLR3 pAb (IMG-516): IHC (P), Fig 4D (mouse eye sections) 2. TLR3-PE mAb (IMG-315D): Flow (Surface) and Flow (Intracellular), Fig 4F [primary human retinal pigment epithelium cell isolates (hRPE)]. TLR3 was expressed on both the cell s
• Desai MM, Gong B, Chan T et al. Differential, type I interferon-mediated autophagic trafficking of hepatitis C virus proteins in mouse liver. Gastroenterology. 2011-08-01 [PMID: 21683701]
• Wu J, Meng Z, Jiang M et al. Toll-like receptor-induced innate immune responses in non-parenchymal liver cells are cell type-specific. Immunology. [PMID: 19922426]
  
  Details:
  Citation using the PE/Cy5 form of this antibody.
• Cho WG, Albuquerque RJ, Kleinman ME et al. Small interfering RNA-induced TLR3 activation inhibits blood and lymphatic vessel growth. Proc Natl Acad Sci U S A. 2009-04-28 [PMID: 19359485]
• Kleinman ME, Yamada K, Takeda A et al. Sequence- and target-independent angiogenesis suppression by siRNA via TLR3. Nature. 2008-04-03 [PMID: 18368052]
• Pawar Rahul D, Patole Prashant S, Ellwart Andreas et al. Ligands to nucleic acid-specific toll-like receptors and the onset of lupus nephritis. J Am Soc Nephrol. 2006-12-01 [PMID: 17082246]
• Patole PS, Pawar RD, Lech M et al. Expression and regulation of Toll-like receptors in lupus-like immune complex glomerulonephritis of MRL-Fas(lpr) mice. Nephrol Dial Transplant. 2006-11-01 [PMID: 16954173] (IHC-Fr, Mouse)
  
  Details:
  For IHC (frozen) on mouse kidney sections (Fig. 5): TLR3 (IMG-516), TLR7 (IMG-581A), TLR9 (IMG-431) and Rabbit IgG isotype control antibody (20304). For Flow (Intacellular) and Flow (Cell surface) on J774 mouse macrophage-like cell line (Fig 6B): TLR7 (IMG-581A), TLR9 (IMG-431) and Rabbit IG isotype control antibody (20304).
• Pawar RD, Patole PS, Zecher D et al. Toll-like receptor-7 modulates immune complex glomerulonephritis. J Am Soc Nephrol. 2006-01-01 [PMID: 16280469] (Flow-CS, Flow Cytometry Control)
  
  Details:
  Antibodies cited: 1. TLR3 (IMG-516) 2. TLR7 (IMG-581) [Flow (intracellular and cell surface), Fig.3: TLR3 (mesengial cells and J774 macrophages), TLR7 (J774 macrophages)]. [IHC-F, Fig.2 (kidneys of MRL1pr/1pr mice)].
• Patole PS, Grone HJ, Segerer S et al. Viral double-stranded RNA aggravates lupus nephritis through Toll-like receptor 3 on glomerular mesangial cells and antigen-presenting cells. J Am Soc Nephrol. 2005-05-01 [PMID: 15772251] (Flow-CS, Flow Cytometry Control)
  
  Details:
  Antibodies cited: 1. TLR3 (IMG-516) [IHC-P, Fig.2 (MRL 1pr/1pr mice kidneys)]. [Flow (cell surface and intracellular), Fig.3 (murine mesangial cell line)].

Review for TLR3 Antibody (NB100-56571) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Other.

reviewed by: Bryan Tinsley IHC Other 09/18/2014

Summary

• Application: Immunohistochemistry
• Species: Other

Bioinformatics

• Gene Symbol: TLR3
• Entrez:
  • 7098(Human)
  • 142980(Mouse)
• Uniprot:
  • O15455()