TLR4 ELISA Kit (Bovine)

ELISA Kit

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TLR4

841

Cooperates with LY96 and CD14 to mediate the innate immune response to bacterial lipopolysaccharide (LPS) (PubMed:17559944). Acts via MYD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Also involved in LPS-independent inflammatory responses triggered by free fatty acids, such as palmitate. In complex with TLR6, promotes sterile inflammation in monocytes/macrophages in response to oxidized low-density lipoprotein (oxLDL) or amyloid-beta 42. In this context, the initial signal is provided by oxLDL- or amyloid-beta 42-binding to CD36. This event induces the formation of a heterodimer of TLR4 and TLR6, which is rapidly internalized and triggers inflammatory response, leading to the NF-kappa-B-dependent production of CXCL1, CXCL2 and CCL9 cytokines, via MYD88 signaling pathway, and CCL5 cytokine, via TICAM1 signaling pathway, as well as IL1B secretion. Binds electronegative LDL (LDL(-)) and mediates the cytokine release induced by LDL(-) (By similarity).

NP_776623.5

This assay has high sensitivity and excellent specificity for detection of Toll Like Receptor 4 (TLR4).No significant cross-reactivity or interference between Toll Like Receptor 4 (TLR4) and analogues was observed.

Component: Amount
Anti-TLR4 Microplate: 96 Wells (12 x 8 Well strips)
TLR4 Lyophilized Standard: 2
100X Biotinylated TLR4 Detector Antibody: 1 x 120 uL
100X Avidin-HRP Conjugate: 1 x 120 uL
Standard Diluent: 1 x 20 mL
Detector Antibody Diluent: 1 x 12 mL
Conjugate Diluent: 1 x 12 mL
30X Wash Buffer: 1 x 20 mL
Stop Solution: 1 x 6 mL
TMB Substrate: 1 x 9 mL

3h

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Toll Like Receptor 4 (TLR4). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Toll Like Receptor 4 (TLR4). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Toll Like Receptor 4 (TLR4), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Toll Like Receptor 4 (TLR4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Toll Like Receptor 4 (TLR4) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Toll Like Receptor 4 (TLR4) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%