Reaction #530971

Reaction equation

Na2CO3

FITC

→

Fluorescein

Solvents

acetone

phosphate

Conditions

Workup

1. 1
   workup.ADDITIONmixed for 3 hrs at room temperature
2. 2
   OtherThe resulting solution was separated two times on Sephadex G-25(M) column (1×15 cm)
3. 3
   Concentrationin concentrated

Procedure

One mg of HIgG (Human IgG) dissolved in 0.4 ml of 0.1 M phosphate buffer pH 7.5, was brought to pH 9.5 with crystalline Na2CO3. A solution (10 μl) of FITC (70 μg) in acetone was added with stirring and mixed for 3 hrs at room temperature. The resulting solution was separated two times on Sephadex G-25(M) column (1×15 cm) equilibrated with 0.05 M phosphate buffer pH 8.0. The eluted FITC-hIgG conjugate solution was 0.58 mg/ml in concentrated and had D/P=5.5 (M/M) as determined by the Wells' nomograph.

Source