Mouse Soluble Toll-like Receptor 4 (sTLR4) ELISA Kit

Tlr4 belongs to the evolutionarily-conserved Toll-like receptor family, whose members are type-1 transmembrane proteins that are involved in innate immunity. Toll-like receptors are characterized by an extracellular leucine-rich repeat domain that functions in ligand recognition and an intracellular toll/interleukin-1 receptor-like domain that is crucial for signal transduction. The receptor encoded by Tlr4 mediates the innate immune response to bacterial lipopolysaccharide, a major component of the outer membrane of Gram-negative bacteria, through synthesis of pro-inflammatory cytokines and chemokines. In addition, this protein can recognize other pathogens from Gram-negative and Gram-positive bacteria as well as viral components. Mice deficient in Tlr4 display a number of immune response-related phenotypes including hyporesponsiveness to bacterial lipopolysaccharide and increased levels of respiratory syncytial virus compared to controls.

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Mouse Soluble Toll-like Receptor 4 (sTLR4) ELISA Kit has high sensitivity and excellent specificity for detection of Mouse TLR4. No significant cross-reactivity or interference between Mouse TLR4 and analogues was observed.

This Mouse Soluble Toll-like Receptor 4 (sTLR4) ELISA Kit employs a two-site sandwich ELISA to quantitate TLR4 in samples. An antibody specific for TLR4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyTLR4 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for TLR4 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of TLR4 bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.