Reacción #666908

Ecuación de reacción

FITC isomer

→

Fluorescein

Disolventes

DMSO

carbonate

Condiciones de reacción

Tratamiento posterior

1. 1
   Otroafter which free (unconjugated) fluorescein is separated
2. 2
   Filtraciónby gel filtration on Sephadex G25 equilibrated in 50 mM sodium phosphate, 0.5M NaCl, and 0.1% azide

Procedimiento

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.

Fuente