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

Participants

Reaction Steps

An IgG fraction of affinity-purified rabbit anti-p19 antibody is fluoresceinated as follows: Thirty micrograms of FITC isomer II (20 μL of a 3.0 mg/mL solution in DMSO) is added to 1 mg of antibody in 125 μL of 0.27M carbonate buffer, pH 9.3. The mixture is incubated for 30 minutes at 37° C., after which free (unconjugated) fluorescein is separated from fluoresceinated antibody by gel filtration on Sephadex G25 equilibrated in 50 mM sodium phosphate, 0.5M NaCl, and 0.1% azide.

Provenance & Evidence