A novel HPLC method for glutathione-insulin transhydrogenase assay using fluorescence labeled insulin.
The activity of rat liver glutathione-insulin transhydrogenase (GIT) was measured by HPLC. The degradation of fluorescein isothiocyanate-I (FITC-I)-labeled insulin is separated into several peaks, which are bound different amount of FITC-I. We selected mono-fluorescein-thiocarbamylated insulin to estimate the decrease of insulin content and it became possible to assay GIT activity. This novel method was time-saving and simple, and this system could utilize instead of previous method.
Saito
N
N
National Institute of Animal Health, Ibaraki, Japan.
Yamanaka
H
H
Nagasawa
S
S
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives*; D007328:Insulin / Q000031:analogs & derivatives* / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008297:Male; D011490:Protein Disulfide Reductase (Glutathione) / Q000032:analysis*; D051381:Rats
D007328:Insulin; C031573:insulin, fluorescein-isothiocyanated-; D011490:Protein Disulfide Reductase (Glutathione); D016650:Fluorescein-5-isothiocyanate