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Datasheet
Monoclonal Antibody to Human TLR3/CD283
Cat.No IMG-315A
Description TLR3/CD283
Format Purified
Unit 0.1 mg
Price(USD) 255.00
MSDS
Catalog No
:
IMG-315A
Contents
:
0.1 mg in 0.2 ml PBS containing 0.05% BSA and 0.05% sodium azide. Sodium azide is highly toxic.
Isotype
:
Mouse IgG1, Kappa
Clone
:
40C1285.6
Purification
:
Protein G Chromatography
Species
:
Dog, Human, Mouse
Host
:
Mouse
Application
Flow (Cell Surface): see Wong et al, 2007 for details Flow (Intracellular): 2-4 ug/ 1x10^6 cells IF/ICC: see Ranjith-Kumar et al, 2007 and IHC (paraffin): (see Furrie et al, 2005) IHC: (see Harii et al, 2005) IP: 2 ug/ 1x10^6 cells Western blot analysis: 1-3 ug/ml Storage Store at 4°C. For long-term storage, store at -20°C
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) and could constitute an important and unrecognized component of innate immunity in humans. The TLR family is a phylogenetically conserved mediator of innate immunity that is essential for microbial recognition (2). TLRs characterized so far activate the MyD88/interleukin-1 receptor-associated kinase (IRAK) signaling pathway. Ten human homologs of TLRs (TLR1-10) have been described (3). TLR3 cDNA codes for a protein with approximate molecular weight of 120 kDa (4). TLR3 has a restricted expression pattern being expressed in dendritic cells (DC) (4-6). TLR3 mRNA expression was detected by in situ hybridization in DC and lymph nodes (6). The expression of TLR3 in a single cell type may indicate a specific role for this molecule in a restricted setting.
Antigen
A synthetic peptide corresponding to amino acids 55-70 (VLNLTHNQLRRLPAAN) of human TLR3 was used as immunogen, Gen Bank no. gb|AAH59372.1| . Human TLR3 is an 870 amino acid protein.
Application Notes
This antibody has tested negative by western blot in mouse intestine cell lysate.
Western blot analysis of TLR3 in lysates from untransfected 293 cells (lane A), 293 cells transfected with human TLR3 cDNA (lane B), and 20 µg/lane human intestine tissue lysate (lane C). IMGENEX's goat anti-mouse HRP conjugate (IMGENEX, 20101) was used as secondary.
Intracellular flow analysis of TLR3 in human monocytes using 0.5 ug of IMG-315A. Shaded histogram represents monocytes without antibody; green represents isotype control (IMGENEX, 20109); red represents anti-TLR3 antibody. Goat anti-mouse IgG1 PE conjugate (BD) was used as secondary.
IHC analysis of TLR3 in (A) human gut lumen (longitudinal section, transverse region) using IMG-315A at 10 ug/ml (Data courtesy Dr. Elizabeth Furrie, University of Dundee) and mouse spleen tissue using isotype control (B) and IMG-315A (C) at 5 mg/ml.
Reference 1. Muzio M, Bosisio D, Polentarutti N, D'amico G, Stoppacciaro A, Mancinelli R, van't Veer C, Penton-Rol G, Ruco LP, Allavena P, Mantovani A. J Immunol 164(11):5998-6004 (2000). 2. Muzio M, Natoli G, Saccani S, Levrero M, and Mantovani A. J. Exp. Med. 187: 2097-2101 (1998). 3. Medzhitov R and Janeway CA. Cell 91: 295-298 (1997). 4. Chuang TH and Ulevitch RJ. Biochim. Biophys. Acta 1518 (1-2): 157-161 (2001) 5. Rock FL., Hardiman G., Timans JC., Kastelein RA. and Bazan JF. Proc. Natl. Acad. Sci. U.S.A. 95 (2), 588-593 (1998). 6. Muzio M, Polentarutti N, Bosisio D, Prahladan MK, Mantovani A. J Leukoc Biol 67(4):450-456 (2000).
Product Citations 1. Biochemical and Functional Analyses of the Human Toll-like Receptor 3 Ectodomain. C.T. Ranjith-Kumar, W Miller, J Xiong, W K. Russell, R Lamb, J Santos, K E. Duffy, L Cleveland, M Park, K Bhardwaj, Z Wu, D H. Russell, R T. Sarisky, M Lamine Mbow, and C. Cheng Kao. J. Biol. Chem., 282, 7668-7678 (2007). Imgenex antibodies cited: 1. TLR3 (IMG-315A) [WB, Fig1A, Fig3B, Fig4D, Fig5D, Fig7B (human TLR3 transfected cells)] 2. TLR3 (IMG-315C) [IF, Fig3C, Fig4E, Fig5E, Fig7C, Fig9D (human transfected cells)]. Antibody specificity validated by WB (IMG-315A) and by IF (IMG-315C) using TLR3 transfected cells. 2. Distinct poly-I: C and virus-activated signaling pathways leading to interferon-b production in hepatocytes. Li K., Zihong Chen, Nobuyuki Kato, Michael Gale ., Jr, and Stanley M. Lemon. J. Biol. Chem., Feb 2005; 10.1074/jbc.M414139200. IMGENEX antibodies cited: 1. TLR3 (IMG-315A) [WB, Fig.6B, (ISG15, ISG56, MxA)]. 3. Toll-like receptors-2, -3 and -4 expression patterns on human colon and their regulation by mucosal-associated bacteria. Furrie E., S. Macfarlane, G. Thompson, and G.T. Macfarlane, Immunology 0 (0), -doi: 10.1111/j.1365-2567.2005.02200.x (2005) Imgenex antibodies cited: 1. TLR3 (IMG-315)[IHC-P, Fig.2 (normal human colon)]. 4. Regulation of the Polymeric Ig Receptor by Signaling through TLRs 3 and 4: Linking Innate and Adaptive Immune Responses. Schneeman T.A., M. E. C. Bruno, H. Schjerven, F-E Johansen, L.Chady, and C. S. Kaetzel. J. Immunol., 175: 376-384 (2005). Imgenex antibodies cited: 1. TLR3 (IMG-315) 2. TLR4 (IMG-417) [IF/ICC, Fig.3 (HT29 cells)]. 5. Subcellular Localization of Toll-Like Receptor 3 in Human Dendritic Cells. Misako Matsumoto, Kenji Funami, Masako Tanabe, Hiroyuki Oshiumi, Masashi Shingai, Yoshiyuki Seto, Akitsugu Yamamoto and Tsukasa Seya. The Journal of Immunology, 171: 3154-3162 (2003). Imgenex antibodies cited: 1. TLR3 (IMG-315A) [IP, Fig. 1 (Ba/F3, HEK293, and HeLa cells)] 6. Recognition of double stranded RNA by human toll like receptor 3 and downstream receptor signaling requires multimerisation and an acidic pH. Odette de Bouteiller, Estelle Merck, Uzma A. Hasan, Sylvain Hubac, Barbara Benguigui, Giorgio Trinchieri, Elizabeth E. M. Bates, and Christophe Caux. J. Biol. Chem., 280: 38133-38145 (2005). Imgenex antibodies cited: 1. TLR3 (IMG-315A) [WB, Fig.1A, 1B, 1C (HEK293 and HEK293T cells)]. 7. Toll-like receptor 4 signaling regulates cytosolic phospholipase A2 activation and lipid generation in LPS-stimulated macrophages. Qi H-Y and J. H. Shelhamer. J. Biol. Chem., Sep 2005; 10.1074/jbc.M509352200. Imgenex antibodies cited: 1. TLR3 (IMG-315) [WB, Fig.3, (RAW264.7)]. 8. Respiratory Syncytial Virus Induces TLR3 Protein and Protein Kinase R, Leading to Increased Double-Stranded RNA Responsiveness in Airway Epithelial Cells. Dayna J. Groskreutz, Martha M. Monick, Linda S. Powers, Timur O. Yarovinsky, Dwight C. Look, and Gary W. Hunninghake. J. Immunol., 176: 1733-1740 (2006). Imgenex antibodies cited: 1. TLR3 (IMG-315) [IF/ICC and Flow (cell surface), Figs. 4, 6 (A549 cells)]. 9. Structural and functional analyses of the human toll-like receptor 3: Role of glycosylation. J. Sun, K. E. Duffy, C. T. Ranjith-Kumar, J. Xiong, R. J. Lamb, J. Santos, H. Masarapu, M. Cunningham, A. Holzenburg, R. T. Sarisky, M. L. Mbow, and C. Kao. J. Biol. Chem., Feb 2006; 10.1074/jbc.M510442200. Imgenex antibodies cited: 1. TLR3 (IMG-315A) [WB, Fig.1C, Fig.2D, Fig.4A (293T cells)]. 10. Toll-Like Receptor 3 signaling evokes a pro inflammatory and proliferative phenotype in human vascular smooth muscle cells. Xin Yang, Vanishree Murthy, Kelly Schultz, Jeffrey B. Tatro, Katherine A. Fitzgerald, and Debbie Beasley. Am J Physiol Heart Circ Physiol, Jun 2006; 10.1152/ajpheart.00252.2006. Imgenex antibodies cited: 1. TLR3 (IMG-315)[WB, used in siRNA assay)]. 11. Toll-like receptor 3 associates with c-Src tyrosine kinase on endosomes to initiate antiviral signaling. Ingvild Bjellmo Johnsen, Thuy Thanh Nguyen, Monika Ringdal, Anne Merete Tryggestad, Oddmund Bakke, Egil Lien, Terje Espevik, Marit W Anthonsen. The EMBO Journal (06 Jul 2006). Imgenex antibodies cited: TLR3 (IMG-315) [IP, Fig.2C (HEK cells)]. 12. Modulation of Neonatal Microbial Recognition: TLR-mediated Innate Immune Responses are Specificallly and Differentially Modulated by Human Milk. LeBouder E, JE Rey-Nores, A-C Raby, M Affolter, K Vidal, CA Thornton, and MO Labeta. J. Immunol., 176:3742-3752 (2006). Imgenex antibodies cited: 1 TLR3 (IMG-315[FC]); 2. TLR5 (IMG-663 [FC]) 13. Effects of Single Nucleotide Polymorphisms on Toll-like Receptor 3 Activity and Expression in Cultured Cells. Ranjith-Kumar CT, W Miller, J Sun, J Xiong, J Santos, I Yarbrough, RJ Lamb, J Mills, KE Duffy, S Hoose, M Cunningham, A Holzenburg, ML Mbow, RT Sarisky, and CC Kao. J Biol Chem www.jbc.org/cgi/doi/10.1074/jbc.M700209200 publication in press (2007). Imgenex antibodies cited: 1. TLR3 (IMG-315A) [WB, Fig 3C, Fig6A (human TLR3 transfected cells)] 2. TLR3 (IMG-315C) [IF (human transfected cells)]. Antibody specificity validated by WB (IMG-315A) and by IF (IMG-315C) using human TLR3 transfected cells. 14. 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). 15. Toll-like receptors in cellular subsets of human tonsil T cells: altered expression during recurrent tonsillitis. Mansson A, M Adner and LO Cardell. Respiratory Resesarch doi:10.1186/1465-9921-7-36. (2006). Imgenex antibodies cited (human tonsils separated into cell subtypes): 1. TLR3 [IMG-315D (Flow-Intracellular), Figs 5 and 6]. 2. TLR5 [IMG-663A (Flow-Intracellular), Fig 6]. 3. TLR9 [IMG-305C (Flow-Intracellular), Fig 4.] 16. Triggering of TLR3 by polyI:C in human corneal epithelial cells to induce inflammatory cytokines. Ueta M., J Hamuro, H. Kiyono, S. Kinoshita., J. BBRC. 331 doi:10.1016/j.bbrc.2005.02.196, 285-294 (2005). Imgenex antibodies cited: 1. TLR3 (IMG-315) [Flow (cell surface), Fig.2 (MRC-5 and HCEC cells), Fig.4C (HCEC, HCFB, MRC-5, and HeLa cells), Fig.6B (HCEC cells)]. 17. Inhibition of neutrophil apoptosis by TLR agonists in whole blood: involvement of the phosphoinositide 3-Kinase/Akt and NF-kB signaling pathways, leading to increased levels of Mcl-1, A1, and phosphorylated bad. François S, J El Benna, M Pham . C Dang, E Pedruzzi, M Gougerot-Pocidalo, C Elbim. J. Immunol 174: 3633-3642 (2005). Imgenex antibodies cited [WB (human polymorphonuclear neutrophils (PMN)), Fig. 5]: 1. TLR2 (IMG-319) 2. TLR3 (IMG-315A) 3. TLR5 (IMG-664) 4. TLR7(IMG-581A) 18. 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)]. 19.
Inhibition of neutrophil apoptosis by TLR agonists in whole blood: involvement of the phosphoinositide 3-kinase/Akt and NF-kB signaling pathways, leading to increased levels of Mcl-1, A1, and phosphorylated Bad. Francois S, J El Benna, PM.C. Dang, E Pedruzzi, MA Gougerot-Pocidalo, and C Elbim., J Immunol, 174: 3633-3642 (2005). IMGENEX antibodies cited: 1. TLR2 (IMG-319) 2. TLR3 (IMG-315) 3. TLR5 (IMG-580) 4. TLR7 (IMG-581) [WB, Fig.5 (PMN cells)].
20. Key differences in TLR3/ply I:C signaling cytokine induction by human primary cells: a phenomenon absent from murine cell systems. Lundberg AM, SK Drexler, C Monaco, LM Williams, SM Sacre, M Feldmann and BM Foxwell. Blood 110:3245-3252 (2007). TLR3-FITC (IMG-315C) used in Flow (Intracellular) and Flow (Cell surface): Fig 1B (human dendritic cells, macrophages, rheumatoid arthritis synovial fibroblasts, and endothelial cells). 21. Identification of toll-like receptor 3 as a potential therapeutic target in a clear cell renal cell carcinoma. Morikawa T, A Sugiyama, H Kume, S Ota, T Kashima, K Tomita, T Kitamura, T Kodama, M Fukayama, H Aburatani. Clinical Cancer Research 13: 5703-5709 (2007). WB: (Human kidney tissues), Fig. 2A. 22. Double-stranded RNA mediates interferon regulatory factor 3 activation and interleukin-6 production by engaging Toll-like receptor 3 in human brain astrocytes. Kim H, E Yang, J Lee, S Kin, J Shin, J Park, S Choi, S Kim, I Choi. Immunology doi: 10.1111/j.1365-2567.2007.02799.x *2008). WB (human brain astrocytes), Fig. 1a,b. 23. STAT3 and STAT5-dependent pathways competitively regulate the pan-differentiation of DC34pos cells into tumor-competent dendritic cells. Cohen P, Koski 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)
24. The cytoplasmic 'linker region' in Toll-like receptor 3 controls receptor localization and signaling. Funami K, M Matsumoto, H Oshiumi, T Akazawa, A Yamamoto, and T Seya. International Immunology 16:1143-1154 (2004). Antibody format cited for WB: IMG-315A (HEK293 cells transfected with various TLR3 plasmid constructs), Fig 3A and 5C,D,F. Note: The IMG-315A antibody is transfected validated in this study. It is also shown to WB TLR3 that has been immunoprecipitated with another TLR3 antibody, Clone TLR3.7 generated in the investigator's laboratory. 25. Down modulation of human TLR3 function by a monoclonal antibody. Duffy KE, RJ Lamb, LR San Mateo, JL Jordan, G Canziani, M Brigham-Burke, J Korteweg, M Cunningham, HS Beck, J Carton, J Giles-Komar, C Duchala, RT Sarisky, ML Mbow. Cellular immunology 248:103-114 (2007). Antibody format cited for WB: IMG-315A (recombinant extracellular domain of human TLR3), Fig 1B. Note: IMG-315A recognized recombinant TLR3 (125 ng) when processed under both denaturing(Fig 1B left) and native (Fig 1B right) conditions. 26.
Antibodies specific for human or murine Toll-like receptors detect canine leukocytes by flow cytometry. Burgener IA and TW Jungi. Veterinary Immunology and Immunopathology 124:184-191 (2008). Imgenex products cited for canine (dog) PBMC subpopulations:
27. Inhibitors of TLR8 reduce TNF production from human rheumatoid synovial membrane cultures. Sacre S, A Lo, B Gregory, R Simmonds, L Williams, M Feldmann, F Brennan, B Foxwell. J Immunol 181: 8002-8009 (2008). Imgenex antibodies cited [Flow (intracellular), human synovial membrane cells, Fig. 4B]: 1. TLR3-FITC (IMG-315C) 2. TLR8-FITC (IMG-321C) 3. TLR9-FITC (IMG-305C). 28.
Sequence- and target-independent angiogenesis suppression by siRNA via TLR3.Kleinman ME, K Yamada, A Takeda, V Chandrasekaran, M Nozaki, JZ Baffi, RJC Albuquerque, S Yamasaki, M Itaya, Y Pan, B Appukuttan, D Gibbs, Z Yang, K Kariko, BK Ambati, TA Wilgus, LA DiPietro, E Sakurai, K Zhang, JR Smith, EW Taylor, J Ambati. Nature 452:591-597 (2008). IMGENEX products cited: 1. IMG-516 (TLR3 pAb): IHC (frozen), Fig S9a (Mouse eye: TLR3 and CD31 signals colocalized at the site of laser injury) 2. IMG-315D (TLR3-PE mAb): Flow (intracellular) and Flow (cell surface), Figs S9f,g (human choroidal endothelial cells) and Fig 26 (human aortic, dermal microvascular, pulmonary microvascular and umbilical vein endothelial cells). TLR3 was detected intracellularly as well as on the cell surface.
29. Extracts of the rat tapeworm, Hymenolepic diminuta, supress macrophage activation in vitro and alleviate chemically induced colitis in mice. Johnston M, A Wang, M Catarino, L Ball, V Phan, J MacDonald, D McKay. Infection & Immunity 78:3 1364-1375 (2010). IMGENEX products cited: 1. TLR4 FITC (IMG-5031A): Flow (cell surface) method described staining on the human monocyte THP-1 cell line, Fig 4A 2. TLR3 PE (IMG-315D): Flow (intracellular) method described on the human monocyte THP-1 cell line. 30. Thyrocytes Express a Functional Toll-like receptor 3 (TLR3): Overexpression Can Be Induced by Viral Infection, Reversed by Phenylmethimazole, and Is Associated with Hashimoto's Autoimmune Thyroiditis. Norikazu Harii, Christopher J Lewis, Vasilly Vasko, Kelly McCall, Uruguaysito Benavides-Peralta, Xiaolu Sun, Matthew D Ringel, Motoyasu Saji, Cesidio Giuliani, Giorgio Napolitano, Douglas J. Goetz, and Leonard D. Kohn. Mol. Endocrinol., 19: 1231-1250 (2005). Imgenex antibodies cited: 1. TLR3 (IMG-315A) [IP, Fig.1B (FRTL-5 cells)], [IHC, Fig.13C and 13D, (normal or Hashimoto's thyroids)]. 31. The Effect of Innate Immunity on Autoimmune Diabetes and the Expression of Toll-Like Receptors on Pancreatic Islets. Li Wen, Jian Peng, Zhenjun Li, and F. Susan Wong. J Immunol 172: 3173-3180 (2004). The antibody was sold by our distributor, BioCarta. Imgenex antibodies cited: 1. TLR3 (IMG-315) [Flow, Fig.4C (J774 cells) and Fig.4D (B6/RIP-B7.1 islet cells)].
Research purposes only.
Not for diagnostic or in vivo use. This product is guaranteed to perform as indicated on the datasheet for one year from the date of purchase.
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