A fluorescence-based assay for indoleamine 2,3-dioxygenase.
A rapid and sensitive fluorescence-based bioassay for determination of indoleamine 2,3-dioxygenase (IDO) activity has been developed. This assay relies on the quantification of the amount of kynurenine produced in the assay medium by fluorescence and complements the standard absorbance and high-performance liquid chromatography (HPLC) assay methods. The fluorescence method has limits of detection similar to those of the standard assay methods. Measured activities of IDO, including in the presence of tryptophan-based inhibitors, were in statistical agreement with the absorbance and HPLC assay methods. The fluorescence-based assay was also suitable for assessment of IDO inhibition by compounds that are incompatible with the absorbance method.
Matin
Azadeh
A
Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, NSW 2109, Australia.
Streete
Isla M
IM
Jamie
Ian M
IM
Truscott
Roger J W
RJ
Jamie
Joanne F
JF
D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000592:standards; D007737:Kynurenine; D012015:Reference Standards; D012972:Sodium Hydroxide; D013050:Spectrometry, Fluorescence / Q000379:methods; D014238:Trichloroacetic Acid
D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase; D007737:Kynurenine; D012972:Sodium Hydroxide; D014238:Trichloroacetic Acid