Bicarbonate Ion → 2-(6-Hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid

HRID 2231183Notes

Reaction equation

HRID 2231183 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].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

3

Reactant 2

HCID 769Bicarbonate IonHCID 18730Fluorescein 5-isothiocyanate

Product 1

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

Procedure

VM-2 antibody (5 mg/ml) was reacted with a tenfold molar excess of fluorescein isothiocyanate immobilized on Celite (Molecular Probes) for 30 minutes in the dark at room temperature in a 0.2M bicarbonate buffer, pH 9.2. Excess fluorescein isothiocyanate was removed by gel chromatography on a Sephadex G50 column in phosphate buffered saline. The fluorophore/protein ratio was calculated from the absorbance of the conjugate at 493 nm and 280 nm and varied between 2.4 to 4.2 from one preparation to another.

Workup

  1. customdark at room temperature
  2. customExcess fluorescein isothiocyanate was removed by gel chromatography on a Sephadex G50 column in phosphate buffered saline
  3. customvaried between 2.4 to 4.2 from one preparation to another

Sources & evidence

ORD record
ord-6bd6e43c9ad3426daf13e23d6aa1f9c7
Source dataset
uspto-grants-1988_11
Patent number
US04782015

Reaction notes

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