The Use of Fluorescein 5′-Isothiocyanate for Studies of Structural and Molecular Mechanisms of Soybean Lipoxygenase

Localization of lipoxygenase activity on the oil bodies and in protoplasts using a novel fluorescence imaging method

“…FITC interacts with lysyl residue on LOX protein surface. Such a fluorescent probe can be used to investigate conformational changes or interactions between the labelled protein and ligands [16]. This probe (FITC), however, is not specific for LOX.…”

“…FITC binding leads to a decrease in enzyme activity, thus making it inappropriate for activity detection. In contrast to FITC, which detects LOX protein, use of H 2 DCFDA provides advantage since it detects LOX activity [16,17]. In recent past, H 2 DCFDA has been used to investigate membrane lipid peroxidation in roots of Arabidopsis [18].…”

9-O-acetylated sialic acids enhance entry of virulentLeishmania donovanipromastigotes into macrophages

“…Briefly, FITC (20 μl, 5 mg/ml) for each mg of protein in dimethyl sulphoxide was incubated in the dark for 2 h at 20°C in borate buffer (50 m m , pH 9·0). Subsequently, the unbound FITC was removed by gradual dialysis with 2–3 changes in borate buffer at 4°C for at least 2 days and the stochiometry of coupling determined before use (García-Barrado et al 1999). The differential presence of 9- O -AcSA on promastigotes was analysed using FITC-Achatinin-H and analysed by a FACS Calibur flow cytometer using CellQuestPro software (Becton Dickinson, Mountain View, CA).…”