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Datasheet

 
Monoclonal Antibody to human TLR2/CD282 PE Conjugate
Cat.No
IMG-416D
Description
TLR2/CD282
Format
PE
Unit
0.1 mg
Price(USD)
330.00
MSDS
View MSDS
Catalog No : IMG-416D
Contents : 100 ug in 200 ul PBS containing 0.05% sodium azide. Sodium azide is highly toxic.
Isotype : Mouse IgG2a, Kappa
Clone : TL2.1
Purification : Protein G Chromatography
Species : Dog, Human
Host : Mouse
Application
Flow (Cell Surface): 0.5-2 ug/10^6 cells
Flow (Intracellular): 0.5-2 ug/10^6 cells
IF/ICC: please see Mempel et. al. (2003) for details
IP: 2-5 ug/ml
Storage
Product is light- and temperature-sensitive.  Store in the dark at 4°C.  Do not freeze.

Background
The Toll-like receptors (TLRs) in mammals comprise a family of transmembrane proteins characterized by multiple copies of leucine rich repeats in the extracellular domain and an 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). TLR2 is differentially expressed in human cells. TLR2 is expressed in tonsils, lymph nodes, and appendices, activated B-cells in germinal centers. CD14+ monocytes expressed the highest level of TLR2 followed by CD15+ granulocytes, and CD19+ B-cells, CD3+ T-cells, and CD56+ NK cells did not express TLR2. The expression of TLR2 in different cell types is regulated by different immune response modifiers. For example, LPS, GM-CSF, IL-1, and IL-10 up regulates TLR2 whereas IL-4, IFN-gamma, and TNF down regulate TLR2 expression in monocytes (4).

Antigen
This antibody was raised by immunizing mice with CHO cells transfected with human TLR2 cDNA (Flo et al, 2000). The hybridoma supernatants were selected by flow cytometry.

Genebank Info (Protein)
NP_003255

 

Flow cytometric analyis of CD14-/CD83+/TLR2+ diffferentiated myeloid dendritic cells (mDCs).  PBMCs were stimulated 5 days with GM-CSF+IL4 prior to the staining.  Cells were stained for surface markers CD14 (IMGENEX, IMG-6350AF647) and CD83-FITC, and TLR2-PE.  CD14- cells were gated and stained with PE-conjugated isotype control (mouse IgG2a) and FITC-conjugated CD83 (left).  CD14- cells were gated and stained with 1 ug PE-conjugated TLR2 (IMGENEX, IMG-416D) and FITC-conjugated CD83 (right).

Cell surface flow analysis of TLR2 in ThP1 cells using IMG-416D at 2 ug/10^6 cells.  Shaded histogram represents ThP1 cells only, green represents isotype control, and red represents TLR2 stained cells.

Cell surface flow analysis of TLR2 in PBMCs (monocytes, live cells gated) using IMG-416D at 0.5 ug/10^6 cells. Shaded histogram represents cells alone; green represents isotype control; red represents TLR2 stained cells.

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Related Products

1. IMG-5019A-1 [Monoclonal Antibody to GAPDH - Loading Control]
2. IMG-5019A-2 [Monoclonal Antibody to GAPDH - Loading Control]
3. IMG-2201 [Pam3CSK4, TLR1 and TLR2 Ligand]
4. IMG-2206 [MALP-2, TLR6/TLR2 ligand]

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. Flo TH, Halaas O, Torp S, Ryan L, Lien E, Dybdahl B, Sundan A, Espevik T. J Leukoc Biol 69(3): 474-481 (2001).
5. Chuang TH and Ulevitch RJ.   Eur Cytokine Netw 11(3): 372-378 (2000).


Product Citations
1. Human Toll-like receptor 2 mediates monocyte activation by Listeria monocytogenes, but not by group B Stretococci or lipopolysaccharide. Flo TH, O Halaas, E Lien, L Ryan, G Teti, DT Golenbock, A Sundan, T Espevik. J Immunology 164:2064-2069 (2000).
Imgenex antibodies cited, TLR2 (IMG-416):
1. Generation of the IMG-416 TL2.1 antibody clone, recognizing a TLR2-associated epitope (Materials and Methods)
2. The IMG-416 antibody is TLR2 transfected validated in Fig 3 [Flow (Cell Surface)], and Fig 4 (IP).
3. Flow (Cell Surface), Fig 3 (human PBMC and TLR2 transfectants)
4. IP, Fig 4 (TLR2 transfectants)
5. FA, Fig 5 [inhibition of Listeria monocytogenes (HKLM) and LPS-induced IL-6 production in transfected cells], and Fig 6 (inhibition of HKLM but not LPS-induced TNF production in human monocytes)

2. Evidence of Toll-like receptor molecules on human platelets. Cognasse F, H Hamzeh, P Chavarin, S Acquart, C Genin, and O Garraud. Immunology and Cell Biology 83:196-198. (2005). The following TLR antibodies were cited in this publication: TLR2-PE (IMG-416D), TLR4-PE (IMG-417D), TLR6 (IMG-304A), TLR8-PE (IMG-321D), TLR9-PE (IMG-305D). Applications: Intracellular Flow Cytomety and Cell Surface Flow Cytometry: Figs 1 and 2. A comparison of staining results, intracellular versus cell surface flow cytometry is shown. Cell type: Human platelets
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. Toll-Like Receptor Expression in Human Keratinocytes: Nuclear Factor B Controlled Gene Activation by Staphylococcus aureus is Toll-Like Receptor 2 But Not Toll-Like Receptor 4 or Platelet Activating Factor Receptor Dependent. Martin Mempel, Verena Voelcker, Gabriele Köllisch, Christian Plank, Roland Rad, Markus Gerhard, Christina Schnopp, Peter Fraunberger, Autar K. Walli, Johannes Ring, Dietrich Abeck, and Markus Ollert. J. Invest. Dermatol., 121: 1389-1396 (2003). Imgenex antibodies cited: 1. TLR2 (IMG-416) 2. TLR4 (IMG-417) [IF/ICC, Fig.2A and 2D (human keratinocytes)].
5. 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)].
6. Highly purified lipoteichoic acid activates neutrophil granulocytes and delays their spontaneous apoptosis via CD14 and TLR2. Sonja Lotz, Eresso Aga, Inga Wilde, Ger van Zandbergen, Thomas Hartung, Werner Solbach, and Tamás Laskay. J. Leukoc. Biol. 75: 467-477 (2004). Imgenex antibodies cited: 1. TLR2 (IMG-416E) 2. TLR4 (IMG-417) [FA, Fig.7C (PMN)].
7. Differential expression of Toll-like receptor 2 in human cells. Flo TH, O Halaas, S Torp, L Ryan, E Lien, B Dybdahl, A Sundan, T Espevik T. J Leukoc Biol 69: 474-481 (2001). Imgenex antibodies cited: TLR2 (IMG-416):
1. Flow (Cell Surface): Figs 1 (whole blood), 5 (granulocytes), and 6 (monocytes)
2. IP: Fig 2 (monocytes)
3. IF/ICC: Fig 3 (monocyte-derived macrophages)

8. 2 TLR2 and TLR4 expression on the immune cells of tuberculous pleural fluid. Prabha C, P Rajashree, D Sulochana. Immunology Letters 117: 26-34 (2008).  Imgenex antibodies cited:
1. TLR2- FITC (IMG-416C): Flow (cell surface): Figs. 1A,B (human CD4+T cells, CD8+T cells, B cells, CD16+56+ cells and monocytes); 2(CD4+T cells); 4A,B (human Treg cells). Flow (intracellular):  Fig. 3A,B (CD4+T cells)
2. TLR4- FITC (IMG-417C).Flow (cell surface): Figs. 1B,C (human CD4+T cells, CD8+T cells, B cells, CD16+56+ cells and monocytes); 2(CD4+T cells); 4A,B (human Treg cells). Flow (intracellular):  Fig. 3A,B (CD4+T cells)
9. Neisseria gonorrhoeae enhances HIV-1 infection of primary resting CD4+T cells through TLR2 activation. Ding J, A Rapista, N Teleshova, G Mosoyan, G Jarvis, M Klotman, T Chang. J Immunol. 184: 2814-2824 (2010). IMGENEX product cited: TLR2/CD282 (IMG-416C). Flow (Cell surface), Fig 6D (human CD4+T cells).
10. 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

11. The probiotic Escherichia coli strain Nissle 1917 induces γς T cell apoptosis via caspase-and FasL-dependent pathways. International Immunology 20:829-840 (2008). Imgenex antibodies cited:
1. IMG-416C (TLR2 FITC) Flow (cell surface), human PBMCs, Fig. 5.
2. IMG-417C (TLR4 FITC) Flow (cell surface), human PBMCs, Fig. 5.

12. Pro-osteogenic phenotype of human aortic value interstitial cells is associated with higher levels of toll-like receptors 2 and 4 and enhanced expression of bone morphogenetic protein 2. Yang X, D Fullerton, X Su, L Ao, J Cleveland, X Meng.  Journal of the American College of Cardiology 53: 491-500 (2009). Imgenex antibodies cited:
1. IMG-416D (TLR2-PE):
WB, Flow (cell surface): human aortic valve interstitial cells, Figs. 1A,B; 2C; 6A.
2. IMG-417D (TLR4-PE): WB, Flow (cell surface): human aortic valve interstitial cells, Figs. 1A,B; 2C; 6A
.
13. Toll-like receptor 2 and NALP2 mediate induction of human Beta-Defensins by Fusobacterium nucleatum in gingical epithelial cells.  Ji S, Shin J E, Kim Y S, Oh J E, Min B M, Choi Y.  Infection and Immunity 77:1044-1052 (2009).  Imgenex antibody cited:
TLR2  [IMG-416C]
Flow (Cell Surface and Intracellular) (HOK-16B cells cocultured with F. nucleatum), Fig. 1d
Flow (Cell Surface) (TLR2-siRNA-transfected HOK-16B cells infected with F. nucleatum), Fig. 4a

14. Expression of mRNA and proteins for Toll-like receptors, associated molecules, defensins and LL-37 by SRIK-NKL, a CD8+ NK/T cell line. Srivastava M D, B I S Srivastava. Leukemia Res 29: 813-820 (2005).
IMGENEX antibodies cited [IF/ICC (SRIK-NKL cells)]:
1.  TLR2 [IMG-416]
2.  TLR4 [IMG-417]
3.  TLR7 [IMG-581]
4.  TLR9 [IMG-305]
15. Innate immune recognition of invasive bacteria accelerates atherosclerosis in apolipoprotein E-deficient mice. Gibson F C III, C Hong, H-H Chou, H Yumoto, J Chen, E Lien, J Wong, C A Genco. Circulation 109: 2801-2806 (2004).
Imgenex antibody cited:  TLR2  [IMG-416]: 
1.  IHC:P (aortic tissue), Fig. 2b 
2.  Flow (Cell Surface) (P gingivalis, mutant P gingivalis), Figs. 3a, 3b

16. Deviation from major codons in the Toll-like receptor genes is associated with low Toll-like receptor expression.  Zhong F, W Cao, E Chan, P N Tay, F F Cahya, H Zhang, J Lu. Immunology, 114: 83 (2005).
Imgenex antibodies cited [Flow (Intracellular) (293T cells and monocytes), Fig. 1]:
1.  TLR2  [IMG-416]
2.  TLR9  [IMG-305]

17. Expression and function of Toll-like receptor 2 in canine blood phagocytes. Bazzocchi C, M Mortarino, S Comazzi, C Bandi, A Franceschi, C Genchi. Veterinary Immunology and Immunopathology 104: 15-19 (2005). Imgenex antibody cited:  TLR2  [IMG-416A]
1.  Flow (whole blood cells from dogs), Fig. 3

18. A dipalmitoylated lipoprotein from Mycoplasma pneumoniae activates NF-kB through TLR1, TLR2, and TLR6.  Shimizu T, Y Kida, K Kuwano.  J. Immunol. 175: 4641-4646 (2005).
Imgenex antibody cited:  TLR2  [IMG-416E]
1.  FA (THP-1 cells), Fig. 2

19. Signal transduction and nuclear responses in Staphylococcus aureus-induced expression of human ß-defensin 3 in skin keratinocytes.  Menzies B E, A Kenoyer. Infect. Immun. 74: 6847-6854 (2006).
Imgenex antibody cited:  TLR2  [IMG-416E]
1.  FA (human keratinocytes), Fig. 2

20. Dysregulation of LPS-induced Toll-like receptor 4-MyD88 complex formation and IL-1 receptor-associated kinase 1 activation inendotoxin-tolerant cells.  Medvedev A E, A Lentschat, L M Wahl, D T Golenbock, S N Vogel.  J Immunol 169: 5209-5216 (2002).
Imgenex antibody cited: TLR2 (IMG-416A):
1.  WB (human monocytes), Fig. 3
2.  Flow (Cell Surface) (human monocytes), Fig. 4

21. Bacterial lipopolysaccharide and IFN-γ induce Toll-like receptor 2 and Toll-like receptor 4 expression in human endothelial cells: Role of NF-κB activation.  Faure E, L Thomas, H Xu, A E Medvedev, O Equils, M Arditi.  J Immunol 166: 2018-2024 (2001).
Imgenex antibody cited:  TLR2 (IMG-416A):
1.  WB (LPS-treated HMEC), Fig. 2a
2.  IHC:P (LPS-stimulated HMEC), Figs. 2b, 2c

22. Human lymphatic endothelial cells express multiple functional TLRs. Pegu A, S Qin, B Junecko, R Nisato, M Pepper, T Reinhart. J Immunol 180: 3399-3405 (2008). The following antibodies were used in flow (cell surface and intracellular) cytometry: TLR2/CD282 PE (IMG-416D), TLR4 (IMG-417D), TLR5 FITC (IMG-663C), TLR6/CD286 FITC (IMG-304C), TLR9/CD289 PE (IMG-305D). Primary human lung microvascular endothelial cells, Fig 1C & 1D.


Note: Data described but not shown for TLR9/CD289 PE (IMG-305D).
23. Differential but direct abolishment of human regulatory T cell suppressive capacity by various TLR2 ligands. Oberg H, T Ly, S Ussat, T Meyer, D Kabelitz, D Wesch. J Immunol 184: 4733-4740 (2010). The following antibodies were used in flow (intracellular) cytometry: TLR2/CD282 PE (IMG-416D) and TLR6/CD286 (IMG-304A). T cells and Tregs were separated from freshly isolated human PBMCs, Fig S3.

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.

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