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

HRID 579726Notes

Reaction equation

HRID 579726 reaction equation
[CH:1]1[C:6](N=C=S)=[CH:5][C:4]2[C:10]([O:12][C:13]3([C:23]4[CH:24]=[CH:25][C:26]([OH:28])=[CH:27][C:22]=4[O:21][C:15]4[CH:16]=[C:17]([OH:20])[CH:18]=[CH:19][C:14]3=4)[C:3]=2[CH:2]=1)=[O:11]>CS(C)=O.C(=O)([O-])[O-]>[CH:1]1[CH:6]=[CH:5][C:4]([C:10]([OH:12])=[O:11])=[C:3]([C:13]2[C:14]3[CH:19]=[CH:18][C:17]([OH:20])=[CH:16][C:15]=3[O:21][C:22]3[C:23]=2[CH:24]=[CH:25][C:26]([CH:27]=3)=[O:28])[CH:2]=1

Participants

4

Reactant 1

HCID 18730Fluorescein 5-isothiocyanate

Product 1

HCID 33832-(6-Hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid

Reagents, catalysts & solvents 2

HCID 679Dimethyl SulfoxideSolventHCID 19660Carbonate IonSolvent

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.

Workup

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

Sources & evidence

ORD record
ord-ae3b3ffc8d0b45aca86b4b090a4bffdc
Source dataset
uspto-grants-1988_06
Patent number
US04749647

Reaction notes

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