Reaction #666908

ord-ae3b3ffc8d0b45aca86b4b090a4bffdc

Reaction equation

O=C1OC2(c3ccc(O)cc3Oc3cc(O)ccc32)c2ccc(N=C=S)cc21
FITC isomer
→
O=C(O)c1ccccc1-c1c2ccc(=O)cc-2oc2cc(O)ccc12
Fluorescein

Reactants

O=C1OC2(c3ccc(O)cc3Oc3cc(O)ccc32)c2ccc(N=C=S)cc21
FITC isomer

Reagents

None

Solvents

CS(C)=O
DMSO
O=C([O-])[O-]
carbonate

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

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

Procedure

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.

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US04749647uspto-grants-1988_06