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KRIBIOLISA™ Nilotinib (TASIGNA™) ELISA

Enzyme Immunoassay for the quantitative determination of Nilotinib in serum, plasma and cell culture supernatant

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Introduction:

Nilotinib (AMN107, trade name Tasigna), in the form of the hydrochloride monohydrate salt, is a smallmolecule tyrosine kinase inhibitor approved for the treatment of imatinib-resistant chronic myelogenous leukemia. Structurally related to imatinib, it was developed based on the structure of the Abl-imatinib complex to address imatinib intolerance and resistance.

Intended Use:

This kit is used for quantitative estimation of Nilotinib in serum, plasma and cell culture supernatant.

Principle:

The Nilotinib ELISA is a competitive immunoassay for the determination of Nilotinib. A constant concentration of Tracer; varying concentrations of unlabeled standard and BCR-ABL Tyrosine Kinase are added in sequence to the uncoated microtitre plate wells and incubated. After 1st incubation, Anti- (BCR-ABL Tyrosine Kinase) Antibody is dispensed to all wells of uncoated Microtitre plate containing the reaction mixture and incubated. After 2nd incubation, all reaction mixtures are transferred to the respective wells of coated microtitre plate and incubated. After washing, Streptavidin HRP conjugate is added and incubated. Captured Tracer is subsequently bound by streptavidin HRP which produces a soluble colored product after addition of TMB substrate. The enzyme reaction is stopped by dispensing of stop solution into the wells.The optical density (OD) of the solution at 450 nm is inversely proportional to the amount of bound Nilotinib molecule present in standards or samples.

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