TLR8 Antibody (44C143) [PE]

Flow Cytometry: TLR8 Antibody (44C143) [PE] [NBP2-24817] - 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.

Flow Cytometry: TLR8 Antibody (44C143) [PE] [NBP2-24817] - Analysis of TLR8 in Ramos cells using 2 ug of NBP2-24817. Shaded histogram represents Ramos cells without antibody; green represents isotype control; red represents anti-TLR8 antibody.

Flow Cytometry: TLR8 Antibody (44C143) [PE] [NBP2-24817] - Intracellular flow cytometric analysis of TLR8 in 10^6 human PBMCs using 0.5 ug of NBP2-24817, 0.25 ug of anti-human CD14, and 0.5 ug of isotype control. Novus's TLR intracellular staining flow assay kit was used to test this product.

Flow Cytometry: TLR8 Antibody (44C143) [PE] [NBP2-24817] - Intracellular analysis of TLR8 in BALB/c mouse splenocytes (lymphocyte gate) using 2 ug of this antibody. Green represents isotype control ; red represents anti-TLR8 antibody.

• Reactivity: Hu, Mu, Rb
• Applications: Flow, IHC
• Clone: 44C143
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: PE

TLR8 Antibody (44C143) [PE] Summary

• Additional Information: Clone 44C143 was used by HLDA to establish CD designation.
• Immunogen: This antibody was developed against a KLH-conjugated synthetic peptide of human TLR8, within amino acids 750-850.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: TLR8
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Flow (Cell Surface)
  • Flow (Intracellular)
  • Flow Cytometry 1ul/1 million cells
  • Immunohistochemistry
• Application Notes: Optimal dilution of this antibody should be experimentally determined.
• Agonist:
  • Imidazoquinoline Resiquimod (R-848), TLR7 and TLR8 ligand NBP2-26231

Reactivity Notes

Rabbit reactivity reported in scientific literature (PMID: 21273544).

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Purity: Protein G purified

Alternate Names for TLR8 Antibody (44C143) [PE]

• CD288 antigen
• CD288
• MGC119599
• MGC119600
• TLR8
• toll-like receptor 8

Background

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

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 TLR8 Antibody (NBP2-24817)(29)

Publications using NBP2-24817

• Deng G, Ge J, Liu C et al. Impaired expression and function of TLR8 in chronic HBV infection and its association with treatment responses during peg-IFN-a-2a antiviral therapy. Clin Res Hepatol Gastroenterol. [PMID: 28236535] (IF/IHC)
  
  Details:
  Citation using the PE form of this antibody.
• Jamin A, Dehoux L, Dossier C et al. Toll-like receptor 3 expression and function in childhood idiopathic nephrotic syndrome Clin Exp Immunol. [PMID: 26123900] (FLOW, Human)
• Zimmermann M, Aguilera FB, Castellucci M et al. Chromatin remodelling and autocrine TNFalpha are required for optimal interleukin-6 expression in activated human neutrophils. Nat Commun. [PMID: 25616107] (FLOW, Human)
  
  Details:
  Citation using the PE form of this antibody.
• Camacho-Sandoval R, Del Rio Estrada PM, Rivero-Arrieta A et al. Differential partial activation phenotype and production of TNF-a by conventional dendritic cells in response to LPS in HIV+ viremic subjects and HIV+ controllers Clin Exp Immunol [PMID: 25130456] (FLOW, Human)
  
  Details:
  Used the PE form of this antibody.
• Green TL, Santos MF, Ejaeidi AA et al. Toll-like receptor (TLR) expression of immune system cells from metastatic breast cancer patients with circulating tumor cells. Exp Mol Pathol. 2014-08-01 [PMID: 24836676] (Flow Cytometry Control, Human)
  
  Details:
  Citation using the Azide Free format of this antibody.
• Wang CH, Eng HL, Lin KH et al. Functional Polymorphisms of TLR8 Are Associated With Hepatitis C Virus Infection Immunology 2013-11-08 [PMID: 24205871] (FLOW, Human)
  
  Details:
  This citation used the PE version of this antibody.
• Abdi J, Mutis T, Garssen J, Redegeld F. Characterization of the Toll-like receptor expression profile in human multiple myeloma cells. PLoS One. 2013-04-08 [PMID: 23593278]
  
  Details:
  Antibodies cited [multiple myeloma (MM) cell lines, human intestine tissue lysate, PBMC, mouse splenocytes] for WB (Figs 2, 3, 4A, 4B, 5, Table 1) and Flow (Intracellular): Figs 4C, 6-8, Table 1: 1. TLR1 (IMG-5012) 2. TLR2 (IMG-416A)3. TLR3 (IMG-315A)4 TL
• Harman AN, Bye CR, Nasr N et al. Identification of lineage relationships and novel markers of blood and skin human dendritic cells. J Immunol. 2013-01-01 [PMID: 23183897]
  
  Details:
  Antibodies cited in Fig 4B for flow cytometric analysis of TLR expression in human monocyte-derived dendritic cells (MDDC), CD14+ monocytes, myeloid DC, and plasmacytoid DC:1. TLR2-FITC, clone T2.1 (IMG-416C): Flow (cell surface)2. TLR4-PE, clone HTA124 (
• Chamberlain ND, Kim SJ, Vila OM et al. Ligation of TLR7 by rheumatoid arthritis synovial fluid single strand RNA induces transcription of TNF-alpha in monocytes. Ann Rheum Dis. 2013-03-01 [PMID: 22730373]
• Berger M, Hsieh CY, Bakele M et al. Neutrophils express distinct RNA receptors in a non-canonical way. J Biol Chem. 2012-06-01 [PMID: 22532562] (Flow-CS, Flow Cytometry Control)
  
  Details:
  TLR8-PE (IMG-321D): Flow (intracellular) + Flow (cell surface), Fig 1A (human neutrophils) & Fig 1B (HL60).2. TLR8 (IMG-321A): IF confocal microscopy, Fig 1C (human neutrophils).3. FITC-conjugated goat anti-rabbit secondary Ab (Cat no 20302), Flow (intracellular) + Flow (cell surface), various Figs.4. FITC-conjugated donkey anti-rat secondary Ab (Cat no 20204), Flow (intracellular) + Flow (cell surface), various Figs.
• Dasgupta G, Chentoufi AA, You S et al. Engagement of TLR2 reverses the suppressor function of conjunctiva CD4+CD25+ regulatory T cells and promotes herpes simplex virus epitope-specific CD4+CD25- effector T cell responses. Invest Ophthalmol Vis Sci. 2011-05-17 [PMID: 21273544] (Flow Cytometry Control, Rabbit)
  
  Details:
  Products cited: Flow (intracellular), Fig 1A ( CD4+CD25+ regulatory T cells purified from rabbit conjunctiva) & Fig 2A (CD11b+ cells isolated from rabbit conjunctiva): TLR2/CD282 PE (IMG-416D), TLR3/CD283 (IMG-315D), TLR4/CD284 (IMG-417D), TLR8/CD288 (IMG
• Schwab N, Zozulya AL, Kieseier BC et al. An imbalance of two functionally and phenotypically different subsets of plasmacytoid dendritic cells characterizes the dysfunctional immune regulation in multiple sclerosis. J Immunol. 2010-05-01 [PMID: 20357264] (Flow - IC, Human)
  
  Details:
  TLR8 (IMG-321A): Flow (intracellular), human pDC's, Fig 1.
• van den Berk LC, Jansen BJ, Siebers-Vermeulen KG et al. Toll-like receptor triggering in cord blood mesenchymal stem cells. J Cell Mol Med. 2009-09-01 [PMID: 20196781] (Human)
  
  Details:
  flow (cell surface) cytometry: TLR5 (IMG-663A), TLR6 (IMG-304), TLR8 (IMG-321). Human mesenchymal stem cells, Fig 1C.
• Biasin M, Piacentini L, Lo Caputo S et al. TLR activation pathways in HIV-1-exposed seronegative individuals. J Immunol. 2010-03-01 [PMID: 20124101] (Flow-CS)
  
  Details:
  TLR8 FITC (IMG-321C): Flow cell surface (CD4+ and CD14+ isolated from PBMC); Fig 3, Fig 4, Table 1.
• Panda A, Qian F, Mohanty S et al. Age-associated decrease in TLR function in primary human dendritic cells predicts influenza vaccine response. J Immunol. 2010-03-01 [PMID: 20100933]
• Clancy RM, Alvarez D, Komissarova E et al. Ro60-associated single-stranded RNA links inflammation with fetal cardiac fibrosis via ligation of TLRs: a novel pathway to autoimmune-associated heart block. J Immunol. 2010-02-15 [PMID: 20089705]
  
  Details:
  Antibodies cited: 1. TLR-7 (IMG-665A): Primary human macrophages derived from PBMCs, Flow (Intracellular): Fig 1A 2. TLR-8 FITC (IMG-321C): Primary human macrophages derived from PBMCs Flow (Intracellular): Fig 1A 3. TLR-7 (IMG-581A): Fetal cardiac fibrob
• 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.
• Pietschmann K, Beetz S, Welte S et al. Toll-like receptor expression and function in subsets of human gammadelta T lymphocytes. Scand J Immunol. 2009-09-01 [PMID: 19703014] (Flow Cytometry Control, Human)
• Sacre SM, Lo A, Gregory B et al. Inhibitors of TLR8 reduce TNF production from human rheumatoid synovial membrane cultures. J Immunol. 2008-12-01 [PMID: 19017992]
• Clark RA, Huang SJ, Murphy GF et al. Human squamous cell carcinomas evade the immune response by down-regulation of vascular E-selectin and recruitment of regulatory T cells. J Exp Med. 2008-09-29 [PMID: 18794336]
  
  Details:
  Antibodies cited: 1. TLR8 (IMG-321A): IF/ICC (human squamous cell carcinoma tumor cells), Fig. 3A. 2. TLR7 (IMG-581A): IF/ICC (human squamous cell carcimona tumor cells), Fig. 3A.
• Cohen PA, Koski GK, Czerniecki BJ et al. STAT3- and STAT5-dependent pathways competitively regulate the pan-differentiation of CD34pos cells into tumor-competent dendritic cells. Blood. 2008-09-01 [PMID: 18577706]
  
  Details:
  Flow (intracellular), mouse bone marrow cells, Fig. 1E: 1. TLR3 FITC (IMG-315C) 2. TLR4 FITC (IMG-417C) 3. TLR7 (IMG-665A) 4. TLR8 FITC (IMG-321C) 5. TLR9 FITC (IMG-305C).
• Zuiderwijk-Sick EA, van der Putten C, Bsibsi M et al. Differentiation of primary adult microglia alters their response to TLR8-mediated activation but not their capacity as APC. Glia. 2007-11-15 [PMID: 17823968] (Human)
  
  Details:
  TLR8/CD288 FITC (IMG-321C). Flow (cell surface): Human macrophage colony stimulation factor (M-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF) differentiated microglia, Fig 4C.
• Kaji M, Tanaka J, Sugita J et al. Ciprofloxacin inhibits lipopolysaccharide-induced toll-like receptor-4 and 8 expression on human monocytes derived from adult and cord blood. Ann Hematol. 2008-03-01 [PMID: 17724596] (Human)
  
  Details:
  TLR8/CD288 (IMG-321A). Human monocytes derived from adult and cord blood, data not shown see paper for more details.
• Murciano C, Villamon E, Yanez A et al. In vitro response to Candida albicans in cultures of whole human blood from young and aged donors. FEMS Immunol Med Microbiol. 2007-11-01 [PMID: 17714490]
  
  Details:
  The following anitbodies were used in flow (cell surface) cytometry: TLR2/CD282 FITC (IMG-416C), TLR4 FITC (IMG-417C), TLR6/CD286 FITC (IMG-304C). Human whole blood from both young and elderly volunteers, Fig 1. Note: The fluorescence mean intensity was m
• Wong CK, Cheung PF, Ip WK et al. Intracellular signaling mechanisms regulating toll-like receptor-mediated activation of eosinophils. Am J Respir Cell Mol Biol. 2007-07-01 [PMID: 17332440]
  
  Details:
  Human blood eosinophils and neutrophils from buffy coat: For WB, Fig. 1A: TLR1 (IMG-5012), TLR5 (IMG-664), TLR6 (IMG-304A), TLR7 (IMG-540), TLR8 (IMG-321A), TLR9 (IMG-305A). For Flow (Intracellular) and Flow (Surface), Fig. 1B: TLR1 (IMG-5021), TLR2 (IMG-416C), TLR3 (IMG-315C), TLR4 (IMG-417C), TLR5 (IMG-663C), TLR6 (IMG-304C), TLR7 (IMG-665A), TLR8 (IMG-321C), TLR9 (IMG-305C).
• Zola H, Swart B, Banham A et al. CD molecules 2006--human cell differentiation molecules J Immunol Methods 2007-01-30 [PMID: 17174972]
  
  Details:
  This citation used the PerCP version of this antibody.
• Xu S, Koldovsky U, Xu M et al High-avidity antitumor T-cell generation by toll receptor 8-primed, myeloid- derived dendritic cells is mediated by IL-12 production. Surgery. 2006-08-01 [PMID: 16904966]
  
  Details:
  Citation using the Azide Free version of this antibody.
• Levy O, Suter EE, Miller RL, Wessels MR. Unique efficacy of Toll-like receptor 8 agonists in activating human neonatal antigen-presenting cells. Blood. 2006-08-15 [PMID: 16638933] (FLOW)
  
  Details:
  TLR8-PE, (IMG-321D): Flow cytometry, Figure 5
• Lee J, Wu CC, Lee KJ et al. Activation of anti-hepatitis C virus responses via Toll-like receptor 7. Proc Natl Acad Sci U S A. 2006-02-07 [PMID: 16446426]

FAQs for TLR8 Antibody (NBP2-24817). (Showing 1 - 1 of 1 FAQs).

1. Do you have any FACS data for your Monoclonal Antibody to human TLR8 (Toll-like receptor 8)/CD288 PE Conjugate - showing that it doesn’t stain TLR8 negative cells? Is there a negative control other than the isotype control data?
   • We don't test the antibody in a TLR8 negative cell model system (we don't have TLR8 knockout models). However, the specificity of the antibody clone (44C143) has been validated in the 2006 HDLA workshop. Here's a link with additional publications citing the antibody clone that are not yet listed on the data sheet: LINK.

Bioinformatics

• Gene Symbol: TLR8