Background
The Toll-like receptor (TLR) family in mammal comprises a family of transmembrane proteins characterized by multiple copies of leucine rich repeats in the extracellular domain and IL-1 receptor motif in the cytoplasmic domain. Like its counterparts in Drosophila, TLRs signal through adaptor molecules (1). The TLR family is a phylogenetically conserved mediator of innate immunity that is essential for microbial recognition (2). Ten human homologs of TLRs (TLR1-10) have been described (3). TLR8 gene contains three exons, two of which have coding function (4). TLR8 cDNA codes for a protein of approximate molecular weight of 120 kDa (4,5).

Antigen
This antibody was developed against a KLH-conjugated synthetic peptide of human TLR8, within amino acids 750-850. It will cross-react with mouse TLR8.

Application Notes

Related Products
1. 10083K [IC-Flow (Intracellular Staining Flow Assay) Kit]
2. IMG-5019A-1 [Monoclonal Antibody to GAPDH - Loading Control]
3. IMG-5019A-2 [Monoclonal Antibody to GAPDH - Loading Control]

Reference
1.  Muzio M, Natoli G, Saccani S, Levrero M, and Mantovani A. J. Exp. Med. 187: 2097-2101 (1998).
2. Medzhitov R and Janeway CA. Cell 91: 295-298 (1997).
3. Chuang TH and Ulevitch RJ. Biochim. Biophys. Acta 1518 (1-2): 157-161 (2001).
4. Du X, Poltorak A, Wei Y, Beutler B. Eur Cytokine Netw 11(3): 362-71 (2000).
5. Chuang TH and Ulevitch RJ.   Eur Cytokine Netw 11(3): 372-378 (2000).

Product Citations
1. Ciprofloxacin inhibits lipopolysaccharide induced toll like receptor 4 and 8 expression on human monocytes derived from adult and cord blood. Ann Hematol 87:229-231 (2008). IMGENEX product cited: TLR8/CD288 (IMG-321A). Human monocytes derived from adult and cord blood, data not shown see paper for more details.
2. Unique efficacy of Toll-like receptor 8 agonists in activating human neonatal antigen-presenting cells. Ofer Levy, Eugenie E Suter, Richard L Miller, and Michael R Wessels. Blood, Apr 2006; 10.1182/blood-2005-12-4821. [TLR8-PE, (IMG-321D): Flow cytometry, Figure 5)
3. Intracellular Signaling Mechanisms Regulating Toll-Like Receptor-Mediated Activation of Eosinophils.  Wong CK, PFY Cheung, WK IP and CWK Lam. Am. J. Respir. Cell Mol. Biol.doi:10.1165/rcmb.2006-0457OC (2007), in press.   Imgenex antibodies cited (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).
4. Activation of anti-hepatitis C virus responses via Toll-like receptor 7. Lee J, CCN Wu, KJ Lee T-H Chuang, K Katakura, Y-T Liu, M Chan, R Tawatao, M Chung, C Shen, HB Cottam, MMC Lai, E Raz and DA Carson. PNAS 103:1828-1833 (2006). Imgenex antibodies cited in this study:  Flow (Intracellular) and Flow (Cell Surface) in human Huh-7 hepatoma cells, Fig. 1A [TLR7 (IMG-665A], [TLR8 (IMG-321C)], and [TLR9 (IMG-305D)];  IHC (paraffin) on Huh-7 cells, normal fibroblasts, HCV-infected liver, and liver carcinoma, Fig 1C-E (TLR7, IMG-665A).  The specificity of the TLR7 antibody (IMG-665A) was validated using TLR7 293 transfected cells in Flow (Intracellular) (Supplemental Data, Fig. 5).  
5. CD molecules 2006 - Human cell differentiation molecules. Zola H, B Swart, A Banham, S Barry, A Beare, A Bensussan, L Boumsell, CD Buckley, H-J Buhring, G Clark, P Engel, D Fox, B0Q Jin, PJ Macardle, F Malavasi, D Mason, H Stockinger, and X Yang. J Immunological Methods 319:1-5 (2007). Imgenex antibodies cited: TLR8 [IMG-321(A-D)] and TLR10 (IMG-386A). Antibody clones 44C143 (TLR8, IMG-321) and 158C114 (TLR10, IMG-386A) confirmed by the HCDM (Human Cell Differentiation Molecules, previously known as HLDA) 2006 Workshop as TLR8/CD288 and TLR10/CD290, respectively. Also see:  http://www.hldaworkshops.org/MoleculeInformation/tabid/54/Default.aspx
6. High-avidity antitumor T-cell generation by toll receptor 8-primed, myeloid-derived dendritic cells is mediated by IL-12 production. Xu S., U. Koldovsky, M. Xu, D. Wang, E. Fitzpatrick, G. Son, G. Koski, and B. J. Czerniecki., Surgery, 140:170-178 (2006). Imgenex antibodies cited: 1. TLR2 (IMG-416) 2. TLR3 (IMG-315) 3. TLR4 (IMG-417) 4. TLR8 (IMG-321) [Flow (cell surface and intracellular), Fig.1A and 1B (human monocytes)].
7. Human squamous cell carcinomas evade the immune response by down-regulation of vascular E-selectin and recruitment of regulatory T cells. Clark R, Huang S, Murphy G, Mollet I, Hijnen, D, Muthukuru M, Schnabacher C, E Vonetta, Miller D, Kim J, Lambert J, Kupper T.  JEM doi: 10.1084/jem.20071190 (2008). Imgenex 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.
8. STAT3 and STAT5-dependent pathways competitively regulate the pan-differentiation of DC34pos cells into tumor competent dendritic cells. Cohen P, Kolski G, Czerniecki B, Bunting K, Fu X, Wang Z, Zhang W, Carter C, Awad M, Distel C, Nagem H, Paustian C, Johnson T, Tisdale J, Shu S. Immunobiology 112:1832-1843 (2008). IMGENEX antibodies cited [Flow (intracellular), mouse bone marrow cells, Supplementary Fig. S4]:
1. TLR3 FITC (IMG-315C)
2. TLR4 FITC (IMG-5031C)
3. TLR7 (IMG-665A)
4. TLR8 FITC (IMG-321C)
5. TLR9 FITC (IMG-305C)
9. An imbalance of two functionally and phenotypically different subsets of plasmacytoid dendritic cells characterizes the dysfunctional immune regulation in multiple sclerosis. Schwab N, A Zozulyia, B Kieseier, K Toyka, H Wiendl. J Immunol 184: 5368-5374 (2010). TLR8 (IMG-321A): Flow (intracellular), human pDC's, Fig 1.
10. Differentiation of primary adult microglia alters their response to TLR8 mediated activation but not their capacity as APC. Zuiderwijk-sick E, C Putten, M Bsibsi, I Deuzing, W Boer, C Deen, I Kondova, L Boven, J Noort, B Hart, S Amor, J Bajramovic. GLIA 55:1589-1600 (2007). IMGENEX product cited: 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.
11. Toll like receptor triggering in cord blood mesenchymal stem cells. Berk L, Bastiaan J, K Vermeulen, M Netea, T Latuhihin, S Bergevoet, R Raymakers,G Kogler, C Fidgor, G Adema, R Torensma. J Cell Mol Med 13: 3415-3426 (2010).
The following antibodies were used in flow (cell surface) cytometry: TLR5 (IMG-663A), TLR6 (IMG-304), TLR8 (IMG-321). Human mesenchymal stem cells, Fig 1C.
12. Toll like receptor expression and function in subsets of human T lymphocytes. Pietschmann K, S Beetz, S Welte, I Martens, J Gruen, H Oberg, D Wesch, D Kabelitz. Scandinavian J Immunol 70: 245-255 (2009).
1. TLR4/CD284 (IMG-417A): Flow (cell surface), Human T cell subsets (V sigma 1 and V sigma 2) isolated from PBMCs, Fig 3.
2. TLR6/CD286 (IMG-304A) & TLR8/CD288 PE (IMG-321D): Flow (intracellular), Human T cell subsets (V sigma 1 and V sigma 2) isolated from PBMCs, Fig 3.
13. In vitro response to Candida albicans in cultures of whole human blood from young and aged donors. Murciano C, E Vilamon, A Yanez, J Murciano, A Mir, D Gozalbo, M Gil. Immunol & Med Microbiol 51: 327-335 (2007). 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 measured in lymphocytes, monocytes, granulocytes (gated by their respective properties in forward and side scatter), Fig 1.
14. 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. Dasgupta G, A Chentoufi, S You, P Falatoonzadeh, L Urbano, A Akhtarmalik, K Nguyen, L Ablabutyan, A Nesburn, L BenMohamed. IOVS doi:10.1167/iovs.10-6522. (2011). IMGENEX 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-321D), TLR9/CD289 (IMG-305D).

FA, Fig 7 (CD4+CD25+ Treg cells): Pam3CSK4 [TLR1 and TLR2 ligand (IMG-2201)], Pam3CSK4 treated and untreated cells were analyzed by Flow (intracellular).

15. Neutrophils express distinct RNA receptors in a non-canonical way. Berger M, C-Y Hsieh, M Bakele, V Marcos, N Riber, M Kormann, L Mays, L Hofer, O Neth, L Vitkov, WD Frautgartner, D von Schweinitz, R Kappler, A Hector, A Weber, D Hart.  JBC 287:19409-19417 (2011). IMGENEX products cited:
1. 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.

16. Ligation of TLR7 by rheumatoid arthritis synovial fluid single strand RNA induces transcription of TNFalpha in monocytes.  Chamberlain ND, S-j Kim, OM Vila, MV Volin, S Volkov, RM Pope, S Arami, AM Mandelin II, S Shahrara. Ann Rheum Dis 72:418-426 (2013).
IMGENEX products: primary human monocytes and macrophages from rheumatoid arthritis patients used in Flow (Intracellular):     
1. TLR7-PE (IMG-665D): Fig 2D
2. TLR8-Alexa647 (IMG-321AF647): Fig 2G
3. IC-Flow (Intracellular Staining Flow Assay) Kit (10083K): Figs 2D, 2G 17. Characterization of the Toll-like receptor expression profile in human multiple myeloma cells. Abdi J, T Mutis, J Garssen, F Redegeld. PLoS ONE 8(4): e60671. doi:10.1371/journal.pone.0060671 (2013). IMGENEX products cited: TLR1 (IMG-5012), TLR2 (IMG-416A), TLR3 (IMG-315A), TLR4 (IMG-5031A), TLR5 (IMG-663A), TLR7 (IMG-581A), TLR8 (IMG-321A) and TLR9 (IMG-305E).