TLR8 Antibody (44C143) [Biotin]

Product Discontinued

TLR8 Antibody (44C143) [Biotin] Summary

• 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:
  • ELISA 1:100-1:2000
  • Flow (Cell Surface)
  • Flow (Intracellular)
• 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. Use in Flow-cell surface reported in scientific literature (PMID: 20124101). Use in Flow-intracellular reported in multiple pieces of scientific literature.
• Agonist:
  • Imidazoquinoline Resiquimod (R-848), TLR7 and TLR8 ligand NBP2-26231
• Readout System:
  • Streptavidin Native Protein NBP2-29367
  • Streptavidin Native Protein NBP2-29370
  • Streptavidin Native Protein NBP2-29371

Reactivity Notes

This antibody is reactive to Human, Mouse

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Purity: Protein G purified

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

• 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 (NBP2-24818)(21)

Publications using NBP2-24818

• 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]
  
  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]
  
  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] (Flow Cytometry Control)
• 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]
• 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.
• 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]

Bioinformatics

• Gene Symbol: TLR8