PathFinder™ IDO1 Inhibitor Mechanism of Action Assay Kit (CAT#: IOK-LX025)

50 reactions

IDO1 His-Tag;
IDO1 Reaction Solution;
IDO1 Assay Buffer;
Reversible Reference Inhibitor (1000x);
Irreversible Reference Inhibitor (1000x);
UV transparent 96-well Reaction plate;
96-well Preincubation Plate;

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.

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.

Gel Packs

Inhibitor and enzyme are preincubated allowing the inhibitor to bind. After preincubation, the mixture is diluted into reaction buffer containing the L-tryptophan (L-Trp) substrate and all necessary coupled reaction components. This allows reversible inhibitors to dissociate, while irreversible inhibitors remain associated with the IDO1 enzyme. IDO1 activity is determined by measuring the absorption of the reaction product at =320 - 325 nm.