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Applications: Useful for the study of IDO1 enzymology, inhibitor screening, and selectivity profiling.
Background: L-tryptophan (L-Trp) is an essential amino acid necessary for proteinsynthesis in mammalian cells and the L-Trp to kynurenine (Kyn) pathway is firmly established asa key regulator of innate and adaptive immunity. Catabolism of L-Trp to Kyn maintains animmunosuppressive microenvironment by starving immune cells of L-Trp and releasingdegradation products of L-Trp that have immunosuppressive functions. Indoleamine 2,3-dioxygenases (IDO1 & IDO2), two of the rate limiting enzymes in this pathway, are upregulatedin many tumors, providing cancer cells with an avenue for immune evasion.
Contraindications: DMSO >0.5%, strong acids or bases, ionic detergents, high salt.Warning: the Fluorescence Solution contains a component that is known to be a skin and eyeirritant, use caution and appropriate personal protective equipment when handling thiscomponent.
Description: The IDO1 Fluorogenic Inhibitor Screening Assay Kit is designed to measure enzyme inhibition of indoleamine 2,3-dioxygenase 1 (IDO1). The IDO1 Fluorogenic Inhibitor Screening Assay Kit is simple to use and detects fluorescence at long wavelengths, which minimizes potential errors due to compound interference. In the assay, the inhibitor and enzyme are added to a sample containing L-Trp substrate. After a 1 hourincubation at room temperature, the fluorescence solution is added and incubated at 37°C for four hours. Activity is measured by reading sample fluorescence at λ=510 nm following excitation of the reaction product at λ=400 nm.
Storage Stability: Stable at least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival
Supplied As: The kit comes in a convenient format, with enough reaction solution and enzyme to perform a total of 96 reactions.
Warnings: Avoid freeze/thaw cycles. The Fluorescence Solution contains a component that is known to be a skin and eye irritant, use caution and appropriate personal protective equipment when handling this component.
Biosafety Level: Not applicable (BSL-1)
References: Seegers, N., et al. J. Biomol. Screen. 2014. 19(9):1266-74.
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