Description: The TL2.1 monoclonal antibody reacts with human Toll-like receptor 2 (TLR2). To date, at least ten 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, a blocking antibody, has been used to study the role of TLR-2 as a pattern recognition receptor in microbial lipoprotein/lipopeptide induced cytokine production from human peripheral blood mononuclear cells. TL2.1 has been reported to immunoprecipitate human TLR2 (~90 kDa) from PBMC and HMEC.
Applications Reported: TL2.1 has been reported for use in flow cytometric analysis and blocking of TLR2-mediated cytokine production. Inhibition of cytokine production has been reported in the literature using Listeria monocytogenes and peptidoglycans. Optimal conditions for the inhibition assays using TL2.1 and specific stimulatory reagents/suppliers have to be evaluated by individual investigators. Neutralization Protocol: (Important note: make sure to eliminate LPS contamination in the culture system) 1) Prepare target cells of interest at the concentration of 2 x 10^6 /mL in 10% FBS-IMDM (or 5% human serum). 2) Pre-incubate the cells with 10-20 µg/mL of TL2.1 or isotype control antibody at room temperature for 30min. 3) Add the stimuli and incubate at 37°C, 5% CO2 for 14 hours. Harvest the supernatants. 4) Follow the protocol of eBioscience Ready-SET-Go ELISA set to detect IL-6 or TNF alpha production.
Applications Tested: The TL2.1 antibody has been tested by flow cytometric analysis of normal human peripheral blood cells. 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 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Storage and handling: Use in a sterile environment.
Filtration: 0.2 µm post-manufacturing filtered.
Purity: Greater than 90%, as determined by SDS-PAGE.
Endotoxin Level: Less than 0.001 ng/µg antibody, as determined by LAL assay.
Aggregation: Less than 10%, as determined by HPLC.
TLR2 is a member of the Toll-like receptor (TLR) family which play a fundamental role in pathogen recognition and activation of innate immunity. TLR proteins act through adaptor molecules such as MyD88 and TIRAP to activate various kinases and transcription factors such as Protein Kinase C (PKC) alpha/beta and NF-kappa-B. TLR2 can form heterodimers with either TLR1 or TLR6, and as a heterodimer, can recognize a variety of bacterial and mycoplasma lipoproteins respectively. TLRs are highly conserved from Drosophila to humans and share structural and functional similarities. The various TLRs exhibit different patterns of expression. TLR2 is expressed most abundantly in peripheral blood leukocytes, and mediates host response to Gram-positive bacteria and yeast via stimulation of NF-kappaB. TLR2 aids in the recognition of pathogen-associated molecular patterns (PAMPs) that are expressed on infectious agents, and mediate the production of cytokines necessary for the development of effective immunity. Ten human homologs of TLRs (TLR1-10) have been described. TLR2 is expressed in human cells such as tonsils, microglia, lymph nodes, and appendices, activated B-cells in germinal centers. CD14+ monocytes express the highest level of TLR2 followed by CD15+ granulocytes, CD19+ B-cells, and CD3+ T-cells. The expression of TLR2 on different cell types are 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: CD282; TLR-2; Toll-like receptor 2; Toll/interleukin-1 receptor-like protein 4
Gene Aliases: CD282; TIL4; TLR2
UniProt ID: (Human) O60603
Entrez Gene ID: (Human) 7097