Fluorescein 5-isothiocyanate → 2-(6-Hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid

Participants

Reaction Steps

Fluoresceinisothiocyanate (FITC) is dissolved in ethanol (5 mg solid/ml). To 2 ml of a 5 mg/ml protein solution from (I), 0.5 ml carbonate buffer (1M NaHCO3 --Na2CO3 buffer pH 9) is added, followed by 50 μl FITC solution. The mixture is shaken well and the free FITC is chromatographically separated from the bound molecules on a Sephadex G50 column using a buffer comprising 10 mM Tris, 1 mM EDTA, 50 mM KCl, pH 7.4. The labeled protein is collected in the void volume.

Provenance & Evidence