L-tryptophan (L-Trp) is an essential amino acid necessary for protein synthesis in mammalian cells and the L-Trp to kynurenine (Kyn) pathway is firmly established as a key regulator of innate and adaptive immunity. Catabolism of L-Trp to Kyn maintains an immunosuppressive microenvironment by starving immune cells of L-Trp and releasing degradation products of L-Trp that have immunosuppressive functions. Indoleamine 2,3- dioxygenases (IDO1 & IDO2), two of the rate limiting enzymes in this pathway, are upregulated in many tumors, providing cancer cells with an avenue for immune evasion.
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 hour incubation 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.
Useful for the study of IDO1 enzymology, inhibitor screening, and selectivity profiling.
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.
IDO1 His-Tag: 140 µg; -80°C IDO1 Fluorogenic Reaction Solution: 3 x 10 ml; -80°C IDO1 Buffer: 3 ml; -80°C Fluorescence Solution: 4 x 1 ml; -80°C Black 384 Well Assay-Plate: 1 Plate sealing film: 1