TLR3 Antibody (40C1285.6) - BSA Free

Novus Biologicals | Catalog # NBP2-24875

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Canine

Cited:

Human, Mouse, Porcine, Bovine, Canine, Ovine

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Block/Neutralize, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, Bioactivity

Cited:

Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Bioassay, Flow Cytometry Control, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 40C1285.6

Format

BSA Free
View all TLR3 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide corresponding to amino acids 55-85 of human TLR3 was used as immunogen.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for TLR3 Antibody (40C1285.6) - BSA Free

Flow Cytometry: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875]

Flow Cytometry: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875]

TLR3-Antibody-40C1285-6-Flow-Cytometry-NBP2-24875-img0014.jpg
Western Blot: TLR3 Antibody (40C1285.6)BSA Free [NBP2-24875]

Western Blot: TLR3 Antibody (40C1285.6)BSA Free [NBP2-24875]

TLR3-Antibody-40C1285-6-Western-Blot-NBP2-24875-img0015.jpg
Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875]

Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875]

Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) [NBP2-24875] - Mouse colon using TLR3 antibody (clone 40C1285.6) at 1:500 dilution with HRP-DAB detection and hematoxylin counterstaining. Intense signal was found in subset of cells at the bases of the crypts.
Flow Cytometry: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875]

Flow Cytometry: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875]

TLR3-Antibody-40C1285-6-Flow-Cytometry-NBP2-24875-img0016.jpg
Western Blot: TLR3 Antibody (40C1285.6)BSA Free [NBP2-24875]

Western Blot: TLR3 Antibody (40C1285.6)BSA Free [NBP2-24875]

Western Blot: TLR3 Antibody (40C1285.6) [NBP2-24875] - Analysis of TLR3 in lysates from A) human intestine and F) ovary using TLR3 antibody at 3 ug/ml. Goat anti-mouse HRP conjugate was used as secondary.
Western Blot: TLR3 Antibody (40C1285.6)BSA Free [NBP2-24875]

Western Blot: TLR3 Antibody (40C1285.6)BSA Free [NBP2-24875]

TLR3-Antibody-40C1285-6-Western-Blot-NBP2-24875-img0017.jpg
Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875]

Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875]

Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) [NBP2-24875] - Tissue section of mouse intestine using TLR3 antibody (clone 40C1285.6) at 1:500 dilution with HRP-DAB detection and hematoxylin counterstaining. The representative image shows a punctate staining of the ER and endosomes in a subset of cells in Peyer's patches (organized lymphoid nodules) in the tested section.
Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875]

Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875]

Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) [NBP2-24875] - Analysis of TLR3 in (A) human gut lumen (longitudinal section, transverse region) using this antibody at 10 ug/ml (Data courtesy Dr. Elizabeth Furrie, University of Dundee) and mouse spleen tissue using isotype control (B) and this antibody (C) at 5 mg/ml.
TLR3 Antibody (40C1285.6) - BSA Free

Immunohistochemistry: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875] -

Immunohistochemistry: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875] - The nasal epithelium expresses TLR3, TLR7, TLR9, RIG-I & MDA-5.Sections of nasal biopsies were incubated with antibodies against TLR3 (A), TLR7 (B), TLR9 (C), RIG-I (D), & MDA-5 (E) & visualized by 3, 3′-diaminobenzidine (brown). In control slides (F), N-series universal negative control reagent was used. All sections were accompanied with a square magnification. All slides were counterstained with haematoxylin (blue). The figure shows one representative biopsy out of four (3 male, 1 female). The arrows indicate positive stained cells. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24886842), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TLR3 Antibody (40C1285.6) - BSA Free

Immunohistochemistry: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875] -

Immunohistochemistry: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875] - TLRs are expressed & functional in chondrocytes.A RT-PCR for TLR3 & 9 expression in primary human & murine chondrocytes. GAPDH served as loading control. B Confocal microscopy of labelled CpG (green) & poly(I:C) (red) uptake by either primary human chondrocytes (scale bar 20 µm) or murine hip caps (scale bar 50 µm (centre) 20 µm (right)). DAPI (blue) was used as counterstaining for nuclei. C IFN alpha ELISA of murine chondrocytes stimulated with ODN (one-way ANOVA:F (2, 9) = 2.21, p = 0.16, N = 4). D IFN alpha ELISA of primary human chondrocytes stimulated with ODN & poly(I:C) (one-way ANOVA:F (3, 11) = 0.75, p = 0.55, N = 4). E IFN beta ELISA of murine chondrocytes stimulated with poly(I:C) or ODN (one-way ANOVA: F (2, 15) = 59.61, p < 0.0001, N = 6). F IFN beta ELISA of primary human chondrocytes stimulated with poly(I:C) or ODN (one-way ANOVA:F (2, 6) = 95.43, p = <0.0001, N = 3). G Staurosporine was used to induce cell death. The release of RNA from human primary chondrocytes after treatment was measured in the culture supernatant using the RiboGreen assay (RM ANOVA: F (4, 12) = 101.2, p < 0.0001, N = 4). Dunnett’s post-hoc test revealed an increase for 200, 500, & 1000 ng/ml staurosporine treatment compared to the untreated control. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35277480), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
TLR3 Antibody (40C1285.6) - BSA Free

Western Blot: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875] -

Western Blot: TLR3 Antibody (40C1285.6) - BSA Free [NBP2-24875] - TLR mRNA & protein expression in airlifted immortal human corneal (HCET) & conjunctival epithelial cells (IOBA-NHC). (a) Averaged fold increase of TLR mRNA in IOBA-NHC cells. The increase was statistically significant for each TLR. (b) Averaged fold increase of TLR mRNA in HCET cells. The increase was statistically significant for each TLR. Cells were airlifted for 10 days. TLR mRNA level was determined by qPCR & compared to submerged cells. The experiment was repeated 3 times. Y error bars represent the standard deviation of the averaged results. (c) Micrographs of a representative western blot showing TLR protein expression in IOBA-NHC cells. Lane 1: submerged-cultured; lane 2: 3 days after airlifting culture; lane 3: 7 days after airlifting culture; lane 4: 10 days after airlifting culture. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was probed as loading control. (d) Averaged TLR protein increase in IOBA-NHC cells 10 days after airlifting culture compared to submerged-cultured cells. X-ray films from 2 independent western blot experiments were scanned in a densitometer, & the results were averaged. The increase was statistically significant for each TLR protein tested. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24976686), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for TLR3 Antibody (40C1285.6) - BSA Free

Application
Recommended Usage

Bioactivity

reported in scientific literature (PMID 28894449)

Block/Neutralize

reported in scientific literature (PMID 28894449)

Flow (Intracellular)

reported in scientific literature (PMID 24836676)

Flow Cytometry

2-4 ug/ 1x10^6 cells

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Western Blot

1-3 ug/ml

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Flow Cytometry

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

Gene Symbol

TLR3

Additional TLR3 Products

Product Documents for TLR3 Antibody (40C1285.6) - BSA Free

Certificate of Analysis

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Product Specific Notices for TLR3 Antibody (40C1285.6) - 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 (40C1285.6) - BSA Free

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Protocols

View specific protocols for TLR3 Antibody (40C1285.6) - BSA Free (NBP2-24875):

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

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