The monoclonal antibody T2.5 recognizes human Toll-like receptor 2 (TLR2). Toll-like receptors (TLR) are highly conserved throughout evolution and have been implicated in the innate defense to many pathogens. At present, ligands for several of the TLR’s, such as TLR2-6,9, have been identified, confirming their role in first line defense against invading microorganism. In mammals, TLRs are identified as type I transmembrane signaling receptors with an extracellular portion containing leucine-rich repeats with pattern recognition capabilities. Pathogen recognition by TLRs provokes rapid activation of innate immunity by inducing proliferation of proinflammatory cytokines and upregulation of costimulatory molecules and eventually toinitiation of adaptive immunity. TLR2 has been identified as a receptor that is central to the innate immune response to lipoproteins of Gram-negative bacteria, several whole Gram-positive bacteria, as well as a receptor for peptidoglycan and lipoteichoic acid and other bacterial cell membrane products. It is suggested that TLR2 is able to recognize such a wide variety of PAMPs (pathogen-specific molecular patterns) by forming heterodimers with other TLRs like e.g. TLR6. TLR2 is essential for recognizing lipopeptides and lipoproteins from several microorganisms and also peptidoglycans derived from gram-positive bacteria. Bacterial species as diverse as mycobacteria, spirochetes, mycoplasma, Staphylococcus aureus, and Streptococcus pneumoniae have all been shown to mediate cellular activation via TLR2.
Flow cytometry, Frozen sections, Functional studies, Immuno assays, Immuno fluorescence, Immuno precipitation
IHC-F: 6µm sections were fixed with aceton. Sections were blocked with goat serum and exposed o/n with T2.5.
FC: 4*104 leukocytes/ml were stained for 30 minutes at 4°C.
FS: mice were injected i.p. with 1 mg T2.5, after 1h incubation mice were challenged;T2.5 5µg/ml was added to cell culture.
IP: 40 µg cleared protein was incubated with 2 µg T2.5 for 1h at 4°C.
IA: T2.5 as a detector.
Dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For functional studies, in vitro dilutions have to be optimized in user’s experimental setting.