TLR8 Antibody (44C143) - Azide and BSA Free

Name: TLR8 Antibody (44C143) - Azide and BSA Free
Brand: Novus Biologicals
SKU: NBP2-77073
Price: 499 USD
Availability: InStock

Novus Biologicals | Catalog # NBP2-77073

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Product Documents

Clone 44C143 was used by HLDA to establish CD designation.

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Product Specifications
Scientific Data
Applications
Flow Cytometry
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse, Rabbit

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, Simple Western, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ kappa Clone # 44C143

Format

Azide and BSA Free

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Product Specifications

Immunogen

This antibody was developed against a KLH-conjugated synthetic peptide of human TLR8, within amino acids 750-850.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁ kappa

Scientific Data Images for TLR8 Antibody (44C143) - Azide and BSA Free

Western Blot: TLR8 Antibody (44C143)Azide and BSA Free [NBP2-77073]

Western Blot: TLR8 Antibody (44C143) - Azide Free [NBP2-77073] - Analysis of TLR8 in human Ramos cell lysate. Goat anti-mouse Ig HRP secondary antibody and PicoTect ECL substrate solution were used for this test. Image from the standard format of this antibody.

Immunohistochemistry-Paraffin: TLR8 Antibody (44C143) - Azide and BSA Free [NBP2-77073]

Immunohistochemistry-Paraffin: TLR8 Antibody (44C143) - Azide Free [NBP2-77073] - Formalin-fixed, paraffin-embedded human spleen stained with TLR8 antibody at 2 ug/ml. Image from the standard format of this antibody.

Flow Cytometry: TLR8 Antibody (44C143) - Azide and BSA Free [NBP2-77073]

Flow Cytometry: TLR8 Antibody (44C143) - Azide Free [NBP2-77073] - Intracellular expression of TLR8 in monocyte-derived macrophages (MDMs). MDMs which were incubated with PE-conjugated anti-TLR8 or isotype antibody control. Intracellular expression was determined by flow cytometry. Representative profiles were similar in four independent experiments (n=4 subjects). Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0104039) licensed under a CC-BY license. Image from the standard format of this antibody.

Western Blot: TLR8 Antibody (44C143)Azide and BSA Free [NBP2-77073]

Western Blot: TLR8 Antibody (44C143) - Azide Free [NBP2-77073] - All the cell lines expressed mRNA for TLR8 (Figure 1) and displayed a strong expression of TLR8 protein (Figure 2). Lysates of human intestinal tissue and mouse splenocytes were used as positive controls. Beta-actin was served as loading control and was used to normalize expression levels between cells. Data are representative for analysis of >=2 independent experiments. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0060671) licensed under a CC-BY license. Image from the standard format of this antibody.

Flow Cytometry: TLR8 Antibody (44C143) - Azide and BSA Free [NBP2-77073]

Flow Cytometry: TLR8 Antibody (44C143) - Azide Free [NBP2-77073] - Expression of TLR8 in Fravel, L363, OPM1, RPMI8226, XG1, and NCI-H929 as determined by flow cytometry. HCMLs were stained using an intracellular staining protocol with TLR8 antibodies followed by relevant secondary fluorescent-conjugated antibodies. Filled histograms (purple) represent the isotype controls and the open histograms (red) indicate TLR8. Data are representative for analysis of >=2 independent experiments. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0060671) licensed under a CC-BY license. Image from the standard format of this antibody.

Flow Cytometry: TLR8 Antibody (44C143) - Azide and BSA Free [NBP2-77073]

Flow Cytometry: TLR8 Antibody (44C143) - Azide Free [NBP2-77073] - HEK cells expressing TLR8 were treated with ssRNA40-Alexa 546 followed by fixation, permeablization and staining for TLR8 using anti-TLR8 Alexa 488 conjugate or isotype control IgG1/Kappa Alexa 488. TLR8 detection is shown along the horizontal axis, and ssRNA40 Alexa 546 detection is shown along the vertical axis. Direct excitation (488 and 561 nm lasers) of both fluorophores (Alexa 488 and Alexa 546) allowed quantitation of cells staining positive for TLR8 (anti-TLR8-Alexa488) and cells that had taken up ssRNA40-Alexa 546. Single-stranded RNA40 binds TLR8 (flow cytometry FRET assay). A representative experiment is shown from three independent experiments with similar results. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0104039) licensed under a CC-BY license. Image from the standard format of this antibody.

Simple Western: TLR8 Antibody (44C143)Azide and BSA Free [NBP2-77073]

Simple Western: TLR8 Antibody (44C143) - Azide Free [NBP2-77073] - Simple Western lane view shows a specific band for TLR8 in 0.5 mg/ml of Ramos (left) and Human Spleen (right) lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. * Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody Image from the standard format of this antibody.

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Applications for TLR8 Antibody (44C143) - Azide and BSA Free

Application

Recommended Usage

Flow (Intracellular)

reported in scientific literature (PMID 24836676)

Flow Cytometry

0.5-1 ug/10^6 cells

Immunohistochemistry-Paraffin

5 ug/ml

Simple Western

20 ug/mL

Western Blot

1-3 ug/ml

Application Notes

Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.

In Simple Western only 10 - 15 ul of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: antibody dilution of 20 ug/mL. Separated by Size-Wes, Sally Sue/Peggy Sue. This antibody is CyTOF ready.

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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

Azide and BSA Free

Preservative

No Preservative

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: TLR8

Toll-like receptor 8 (TLR8) is a member of the TLR family of receptors that play a role in innate immune system activation and the recognition of pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs) (1,2). TLRs are type I membrane receptors that can be expressed on either the cell surface or internally, on endosomes (1,2). TLR8 is an endosomal receptor and is activated by pathogenic single stranded (ss) RNA (1-3). TLR8 is located on the X chromosome and is expressed mostly in monocytes/macrophages, neutrophils, and myeloid dendritic cells (1-3). Structurally, TLR8 consists of an extracellular domain, a cysteine-rich region, and transmembrane domain, and a Toll/Interleukin-1 receptor homology (TIR) domain (3,4). The extracellular domain contains a N-terminal leucine rich repeat (LRRNT) and a C-terminal LRR (LRRCT) which have 26 LRRs between them, each approximately 20-30 amino acids (aa), and a Z-loop between LRR14 and LRR15 (3). The primary isoform of the human TLR8 is synthesized as a protein 1041 aa in length with a theoretical molecular weight of ~120 kDa (4).

TLR8 is highly similar to TLR7 and both pathways are mediated by the adapter protein MyD88 to signal through IFN regulatory factor 7 (IRF7) and nuclear factor (NF)-kappaB (1-3,5). However, TLR7 recognizes guanosine and GU-rich ssRNA, while TLR8 recognizes uridine and AU-rich sequences (2,5). TLR7/TLR8 agonists, including derivatives of the immunostimulatory imiquimod, have been shown to be a promising cancer therapy capable of providing anticancer signals to antigen presenting cells (APCs), with many agonists being tested in both pre-clinical and clinical trials (6). Similarly, studies suggest that agonists for TLR8, in combination with other individual TLR agonists and antagonists, may also be useful for treating inflammatory allergic diseases, such as allergic rhinitis (7).

References

1. Sakaniwa, K., & Shimizu, T. (2020). Targeting the innate immune receptor TLR8 using small-molecule agents. Acta crystallographica. Section D, Structural biology, 76(Pt 7). https://doi.org/10.1107/S2059798320006518

2. Cervantes, J. L., Weinerman, B., Basole, C., & Salazar, J. C. (2012). TLR8: the forgotten relative revindicated. Cellular & molecular immunology. https://doi.org/10.1038/cmi.2012.38

3. Ohto, U., Tanji, H., & Shimizu, T. (2014). Structure and function of toll-like receptor 8. Microbes and infection. https://doi.org/10.1016/j.micinf.2014.01.007

4. Uniprot (Q9NR97)

5. Jannuzzi, G. P., de Almeida, J., Paulo, L., de Almeida, S. R., & Ferreira, K. S. (2020). Intracellular PRRs Activation in Targeting the Immune Response Against Fungal Infections. Frontiers in cellular and infection microbiology. https://doi.org/10.3389/fcimb.2020.591970

6. Frega, G., Wu, Q., Le Naour, J., Vacchelli, E., Galluzzi, L., Kroemer, G., & Kepp, O. (2020). Trial Watch: experimental TLR7/TLR8 agonists for oncological indications. Oncoimmunology. https://doi.org/10.1080/2162402X.2020.1796002

7. Golshiri-Isfahani, A., Amizadeh, M., & Arababadi, M. K. (2018). The roles of toll like receptor 3, 7 and 8 in allergic rhinitis pathogenesis. Allergologia et immunopathologia. https://doi.org/10.1016/j.aller.2017.09.026

Long Name

Toll-like Receptor 8

Alternate Names

CD288

Gene Symbol

TLR8

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Product Specific Notices for TLR8 Antibody (44C143) - Azide and 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.