Description
Introduction:
Everolimus is an mTOR inhibitor that binds with high affinity to the FK506 binding protein-12 (FKBP-12), thereby forming a drug complex that inhibits the activation of mTOR. This inhibition reduces the activity of effectors downstream, which leads to a blockage in the progression of cells from G1 into S phase, and subsequently inducing cell growth arrest and apoptosis. Everolimus also inhibits the expression of hypoxia-inducible factor, leading to a decrease in the expression of vascular endothelial growth factor. The result of everolimus inhibition of mTOR is a reduction in cell proliferation, angiogenesis, and glucose uptake.
Intended Use:
The KRIBIOLISA Everolimus (AFINITOR) ELISA is used as an analytical tool for quantitative determination of Everolimus (AFINITOR) in serum, plasma and cell culture supernatant.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Everolimus (AFINITOR) are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Everolimus (AFINITOR) present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti-Everolimus (AFINITOR) antibody is pipetted and incubated. After washing microwells in order to remove any non-specific binding, the ready to use substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Everolimus (AFINITOR) in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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