Staining of normal human peripheral blood cells with 0.5 µg of Mouse IgG2a K Isotype Control Purified (cat. 14-4724) (open histogram) or 1.0 µg of Anti-Human CD282 (TLR2) Purified (filled histogram) followed by Anti-Mouse IgG FITC (cat. 11-4011). Cells in the monocyte gate were used for analysis.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The TL2.3 monoclonal antibody reacts with human Toll-like receptor 2 (TLR2). To date, at least twelve members of the Toll family have been identified in human. This family of type I transmembrane proteins is characterized by an extracellular domain with leucine-rich repeats and a cytoplasmic domain with homology to the type I IL-1 receptor. Two of these receptors, TLR2 and TLR4, are pattern recognition receptors and signaling molecules in response to bacterial lipoproteins and have been implicated in innate immunity and inflammation. TLR2 is expressed by peripheral blood monocytes and is responsible for distinguishing different pathogens. TL2.1 and TL2.3, reported as blocking antibodies, have been used to study the role of TLR2 as a pattern recognition receptor in microbial lipoprotein/lipopeptide induced cytokine production from human peripheral blood mononuclear cells. TL2.3 has been reported to recognize a different epitope than TL2.1 and to work better for detection of TLR2 by immunohistochemistry. TL2.1 and its directly fluorochrome conjugates are more efficient for detection of human TLR2 by flow cytometric analysis.
Applications Reported
For research use only, not for diagnostic or therapeutic use. TL2.3 has been reported for use in flow cytometric analysis, and immunohistochemical staining of frozen tissue sections. The reported applicability of TL2.1 and TL2.3 for immunoblotting, histochemistry and blocking assays may require high expression level of TLR2 and has not been confirmed in our tests.
Applications Tested
The TL2.3 antibody has been tested by flow cytometric analysis of human peripheral blood leukocytes. This can be used at less than or equal to 1 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Flo, T. H., O. Halaas, et al. (2000). “Human toll-like receptor 2 mediates monocyte activation by Listeria monocytogenes, but not by group B streptococci or lipopolysaccharide.” J Immunol 164(4): 2064-9.
Lien, E., T. J. Sellati, et al. (1999). “Toll-like receptor 2 functions as a pattern recognition receptor for diverse bacterial products.” J Biol Chem 274(47): 33419-25.
Faure, E., L. Thomas, et al. (2001). “Bacterial lipopolysaccharide and IFN-gamma induce Toll-like receptor 2 and Toll-like receptor 4 expression in human endothelial cells: role of NF-kappaB activation.” J. Immunol 166(3): 2018-24.
