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Datasheet

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Monoclonal Antibody to TLR9/CD289 (Clone 26C593.2) Alexa Fluor® 488 Conjugate
Cat.No
IMG-305AF488
Description
TLR9/CD289
Unit
0.1 mg
Price(USD)
330.00
MSDS
View MSDS
Catalog No : IMG-305AF488
Contents : 100 ug in 200 ul PBS containing 0.05% BSA and 0.05% sodium azide. Sodium azide is highly toxic.
Isotype : Mouse IgG1, Kappa
Clone : 26C593.2
Purification : Protein G Chromatography
Species : Dog, Horse, Human, Mouse, Rat, Rhesus Monkey
Host : Mouse
Application
Confocal Microscopy: see Greene et. al. (2005)
Flow (Cell Surface): see Lee et al. (2006)
Flow (Intracellular): 0.5-1 ug/10^6 cells
IF/ICC: see Greene et. al. (2005)
Storage
Store at 4°C, stable for 6 months. DO NOT FREEZE; MATERIAL IS LIGHT-SENSITIVE.
Recommended Positive Control : Ramos cells

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). By using a BLAST search, Hemmi et al., 2000 (5) have identified and subsequently isolated a cDNA coding for TLR9. Gene knockout experiments suggest that TLR9 acts as a receptor for unmethylated CpG dinucleotides in the bacterial DNA (4). Human and mouse TLR9 share an overall amino-acid identity of 75.5%. TLR9 is highly expressed in spleen.

Antigen
This antibody was developed against KLH-conjugated synthetic peptide corresponding to amino acids 268-284 (CPRHFPQLHPDTFSHLS) of human TLR9 isoform A (Genbank accession no. AAF78037).

Genebank Info (Protein)
NP_059138

 

Intracellular flow analysis of TLR9 in Ramos cells using 0.5 ug of IMG-305AF488. Shaded histogram represents Ramos cells without antibody; green represents isotype control (IMGENEX, 20108); red represents anti-TLR9 antibody.  IMGENEX's TLR intracellular flow kit (IMGENEX, 10098K) was used for this test.

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Related Products

1. IMG-305B [Monoclonal Antibody to TLR9/CD289 (Clone 26C593.2) Biotin Conjugate]
2. IMG-305D [Monoclonal Antibody to TLR9/CD289 (Clone 26C593.2)]
3. IMG-305C [Monoclonal Antibody to TLR9/CD289 (Clone 26C593.2)]
4. 10083K [IC-Flow (Intracellular Staining Flow Assay) Kit]

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. Takeuchi O, Kawai T, Sanjo H, Copeland NG, Gilbert DJ, Jenkins NA, Takeda K, and Akira S. Gene 231: 59-65 (1999).
5. Hemmi H, Takeuchi O, Kawai T, Kaisho T, Sato S, Sanjo H, Matsumoto M, Hoshino K, Wagner H, Takeda K, and Shizo A. Nature 408: 740-745 (2000).


Product Citations
1. Expression of mRNA and proteins for toll-like receptors, associated molecules, defensins and LL-37 by SRIK-NKL, a CD8+ NK/T cell line. Maya D. Srivastava and B.I.S. Srivastava. Leukemia Res., 29: 813-820 (2005). (IMG-305A)
2. TLR-Induced Inflammation in Cystic Fibrosis and Non-Cystic Fibrosis Airway Epithelial Cells. Catherine M. Greene, Tomás P. Carroll, Stephen G. J. Smith, Clifford C. Taggart, James Devaney, Siobhan Griffin, Shane J. O’Neill, and Noel G. McElvaney. J. Immunol., 174: 1638-1646 (2005). Imgenex antibodies cited: 1. TLR9 (IMG-305) [Confocal Microscopy, Fig.1A and 1B (CFTE29o- and 16HBE14o- cells)].
3. Cytoplasmic domain-mediated dimerizations of toll-like receptor 4 observed by lactamase enzyme fragment complementation. Hyun-Ku Lee, Stefan Dunzendorfer, and Peter S. Tobias.  J. Biol. Chem., 279: 10564-10574 (2004). (IMG-305A)
4. Evidence of Toll-like receptor molecules on human platelets. Fabrice Cognasse, Hind Hamzeh, Patricia Chavarin, Sophie Acquart, Christian Genin and Olivier Garraud. Immunology and Cell Biology. Volume 83 Issue 2 Page 196 - April 2005. doi:10.1111/j.1440-1711.2005.01314.x (IMG-305A)
5. The Unc93b1 mutation 3d disrupts exogenous antigen presentation and signaling via Toll-like receptors 3, 7 and 9. Koichi Tabeta, Kasper Hoebe, Edith M Janssen, Xin Du, Philippe Georgel, Karine Crozat, Suzanne Mudd, Navjiwan Mann, Sosathya Sovath, Jason Goode, Louis Shamel, Anat A, et al. Nature Immunology 7, 156-164 (2006). Imgenex antibodies cited: 1. TLR9 (IMG-305) [IF/ICC, Fig.5g-i (primary mouse macrophages)]. The specificity of the TLR9 antibody is demonstrated using primary mouse macrophages from wildtype (5g) and TLR9 knockout (5i) mice. 

6. Novel Toll-Like Receptor 9 Agonist Induces Epidermal Growth Factor Receptor (EGFR) Inhibition and Synergistic Antitumor Activity with EGFR Inhibitors. Vincenzo Damiano, Rosa Caputo, Roberto Bianco, Francesco P. D'Armiento, Antonio Leonardi, Sabino De Placido, A. Raffaele Bianco, Sudhir Agrawal, Fortunato Ciardiello, and Giampaolo Tortora. Clin. Cancer Res., 12: 577-583 (2006). (IMG-305A)
7. 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).
8. Toll-like receptor 9 expression in murine and human adrenal glands and possible implications during inflammation. Tran N, A Koch, R Berkels, O Boehm, PA Zacharowski, G Baumgarten, P Knuefermann, M Schott, W Kanczkowski, SR Bornstein, SL LIghtman, and K Zacharowski. J Clin Endocrin Metab. Doi:10.1210/jc.2006-2697 In press (2007). Imgenex antibodies cited (mouse spleen and adrenal tisse, mouse RAW264.7 macrophage cell line, human adrenal tissue: TLR9 [(IMG-305A) IHC (paraffin): Figs 1C-D, F and 6; IF/ICC: Figs 1E and 5]; TLR9 [(IMG-431) WB: Figs 1B, 3C, and 5G].  The TLR9 IMG-431 antibody was validated by WB using adrenal gland tissue from TLR9-/- and wt mice (Fig 1B).    
9. Deoxycytidyl-deoxyguanosine oligonucleotide classes A, B, and C induce distinct cytokine gene expression patterns in Rhesus monkey peripheral blood mononuclear cells and distinct alpha interferon responses in TLR9-expressing Rhesus monkey plasmacytoid dendritic cells. Abel K, Y Wang, L Fritts, E Sanchez, E Chung, P Fitzgerald-Bocarsly, AM Krieg, and CJ Miller. Clinical and Diagnostic Laboratory Immunology 12:606-621 (2005).  Imgenex antibodies cited: IMG-305C [Flow (Intracellular), Fig. 5] on Rhesus monkey spleen cell suspensions and PBMC.
10. 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.] .
11. Maintenance of colonic homeostasis by distinctive apical TLR9 signaling in intestinal epithelial cells. Lee J., J-H Mo, K. Katakura, I. Alkalay, A. N. Rucker, Y-T. Liu, H-K. Lee, C. Shen, G. Cojocaru, S. Shenouda, M. Kagnoff, L. Eckmann, Y. Ben-Neriah, and E. Raz., Nature Cell Biol., 8,doi:10.1038/ncb1500 (2006). Imgenex antibodies cited: 1. TLR9 (IMG-305) [Confocal Microscopy, Fig.2 (intestinal epithelial cells)]. [Flow (cell surface), Fig.S1 (293-HEK TLR9 transfected cells) and Fig.S2 (Caco-2 intestinal epithelial cells)], TLR9 mAb was transfected validated by flow (cell surface) in Fig. S1. [WB and IP, Fig S1 (293-HEK TLR9 transfected cells and Caco-2 intestinal epithelial cells), TLR9 mAb was transfected validated by IP/WB.

12. Role of pathogenic auto-antibody production by Toll-like receptor 9 of B cells in active systemic lupus erythematosus. Nakano S, S Morimoto, J Suzuki, K Nozawa, H Amano, Y Tokano, Y Takasaki. Rheumatology 47: 145-149 (2008). Imgenex antibody cited: IMG-305C (TLR9-FITC) for Flow (Intracellular),  human peripheral blood B cells from normal and SLE patients, Figs 1C, 2, 3.
13. Induction of pro-inflammatory programs in enteroendocrine cells by the Toll-like receptor agonists flagellin and bacterial LPS. Selleri S, Palazzo M, Deola S, Wang E, Balsari A, Marincola F, Rumio C.  International Immunology doi:10.1093/intimm/dxn055 (2008). TLR9-FITC (IMG-305C): Flow (intracellular), Human colon neuroendocrine LCC-18 cell line: Fig. 4d.
14.

Molecular cloning and characterization of equine Toll-like receptor 9. Zhang YW, EG Davis, F Blecha, MJ Wilkerson. Veterinary Immunology and Immunopathology 124: 208-219 (2008)..

Imgenex Products cited for Flow (Cell Surface) and Flow (Intracellular):.

IMG-305C (TLR9-FITC) and IMG-305D (TLR9-PE): Fig 2B (HEK293 and equine TLR9 transfected cells), Fig 4 (equine leukocytes), Fig 5 (equine PHA-stimulated PBMC), Fig 6 (equine leukocyte subsets). .

Notes: TLR9 expression was found to be predominantly intracellular. HEK293 cells express endogenous TLR9. TLR9 expression increased in equine TLR9 transiently transfected HEK293 cells. TLR9 expression increased in PBMC cells following PHA stimulation.  

15.
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)

16.
Antibodies specific for human or murine Toll-like receptors detect canine leukocytes by flow cytometry. Burgener IA and TW Jungi. Veterinary Immunology and Immunopathology124:184-191 (2008). Imgenex products cited for canine (dog) PBMC subpopulations:
1. 10083K [IC-Flow (Intracellular Staining Flow Assay) Kit
2. IMG-416A (TLR2), Flow (Cell Surface), Fig 1 and Tables 1 & 2
3. IMG-315A (TLR3), Flow (Intracellular), Tables 1 & 2
4. IMG-417A (TLR4), Flow (Intracellular), Fig 1 and Tables 1 & 2
5. IMG-663A (TLR5), Flow (Cell Surface), Fig 1 and Tables 1 & 2
6. IMG-664A (TLR5), Flow (Cell Surface), Tables 1 & 2
7. IMG-305A (TLR9), Flow (Intracellular), Fig 2 and Tables 1 & 2

17.

Stimulation of iNOS expression and apoptosis resistance in B-cell chronic lymphocytic leukemia (B-CLL) cells through engagement of Toll-like receptor 7 (TLR-7) and NF-kB activation. Hammadi A, Billard C, A-M Faussat, J-P Kolb. Nitric Oxide 19:138-145 (2008).Imgenex products cited (B-CLL cells from a leukemia patient):

1.  IMG-417E (TLR4-FITC, clone HTA125): Flow (Intracellular), Fig 1: TLR4 was undetectable in the B-CLL cells.

2. IMG-305C (TLR9-FITC, clone 26C593.2): Flow (intracellular), Fig 1: TLR9 was detected in the BCLL cells

3. IMK-501 (Phospho-IkBa ActivELISA Kit): Fig 6B, ELISA data shows increased of IkBa phosphoryation following resiquimod treatment which stimulates NF-kB activation. 
 
Note: No expression of TLR-4 or TLR-9 was detected at the surface of  B-CLL cells by Flow (cell surface).  

18. Deoxynucleic acids from Cryptococcus neoformans activate myeloid dendritic cells via a TLR9-dependent pathway. Nakamura K, A Miyazato, G Xiao, M Hatta, K Inden, T Aoyagi, K Shiratori, K Takeda, S Akira, S Saijo, Y Iwakura, Y Adachi, N Ohno, K Suzuki, J Fujita, M Kakau, and K Kawakami.  J Immunol 180:4067-4074 (2008). Imgenex antibody used: TLR9-FITC (IMG-305C) for IF/ICC (intracellular confocal microscopy) in the following figures:
1. Fig 8a: primary mouse bone marrow-derived myeloid dendritic cells (BM-DC) treated with Cn-DNA and CpG-Rhodamine and analyzed over a time course
2. Fig 8b: BM-DC and RAW264.1 cells treated with Cn-DNA  
Note: intracellular redistribution of TLR9 was observed in treated cells

19. Members of the Toll-like Receptor Family of Innate Immunity Pattern-Recognition Receptors Are Abundant in the Male Rat Reproductive Tract. Palladino MA., TA Johnson, R Gupta, JL Chapman, and P Ojha. Biol Reproduction., in press, DOI: 10.1095/biolreprod.106.059410 (2007). IMGENEX antibodies cited: The following antibodies were used for WB in Fig. 2 using rat, testis, epididymis, and vas deferens: TLR 1 (IMG-5012), TLR2 (IMG-545), TLR5 (IMG-580), TLR6 (IMG-527), TLR7 (IMG-581) TLR9 (IMG-305A), TLR10 (IMG-5255A), TLR11 (IMG-5034) and MYD88 (IMG-178).

Research purposes only. Not for diagnostic or in vivo use. This product is guaranteed to perform as indicated on the datasheet for six months from the date of purchase.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc. IMGENEX sells Alexa Fluor® dye conjugated antibodies under license from Molecular Probes, Inc. for research use only.

Related Products

Related Product(s) of the Catalog # IMG-305AF488
Cat. No Description Format Species Appl. Qty Price(USD)
IMG-305B TLR9/CD289 Biotin D, H, H, M, R, R ELISA 0.1 mg 275.00 Add to Cart
IMG-305D TLR9/CD289 PE D, H, H, M, R, R Confocal Microscopy, Flow (Cell Surface), Flow (Intracellular), IHC (frozen), IHC (paraffin) 0.1 mg 345.00 Add to Cart
IMG-305C TLR9/CD289 FITC D, H, H, M, R, R Confocal Microscopy, Flow (Cell Surface), Flow (Intracellular), IF/ICC, IHC (frozen), IHC (paraffin), WB 0.1 mg 305.00 Add to Cart
Cat. No Description Species Qty Price(USD)
10083K IC-Flow (Intracellular Staining Flow Assay) Kit 1 kit 145.00 Add to Cart

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