Sodium Chloride → 2-(6-Hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid

HRID 2422999Notes

Reaction equation

HRID 2422999 reaction equation
P([O-])([O-])([O-])=O.[Na+].[Na+].[Na+].[Na+].[Cl-].[CH:11]1[C:16](N=C=S)=[CH:15][C:14]2[C:20]([O:22][C:23]3([C:33]4[CH:34]=[CH:35][C:36]([OH:38])=[CH:37][C:32]=4[O:31][C:25]4[CH:26]=[C:27]([OH:30])[CH:28]=[CH:29][C:24]3=4)[C:13]=2[CH:12]=1)=[O:21]>CS(C)=O>[CH:11]1[CH:16]=[CH:15][C:14]([C:20]([OH:22])=[O:21])=[C:13]([C:23]2[C:24]3[CH:29]=[CH:28][C:27]([OH:30])=[CH:26][C:25]=3[O:31][C:32]3[C:33]=2[CH:34]=[CH:35][C:36]([CH:37]=3)=[O:38])[CH:12]=1

Participants

5

Reactant 3

HCID 5234Sodium ChlorideHCID 18730Fluorescein 5-isothiocyanateHCID 24243Trisodium Phosphate

Product 1

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

Reagents, catalysts & solvents 1

HCID 679Dimethyl SulfoxideSolvent

Procedure

Recombinant purified proteins were labelled with fluorescein-isothiocyanate (FITC, Molecular Probes F143). Reaction was carried out by adding proteins to 10 mM sodium phosphate, 50 mM NaCl buffer pH 7 at a final protein concentration of 1 mg/ml. Labelling reaction was started by adding FITC dissolved in DMSO at a concentration of 20 mg/ml to reach 20:1 molar ratio with respect to the protein concentration. The mixture was left to react at room temperature for an hour. Labelled protein was separated from the free fluorescein on a PD-10™ column (Pharmacia) prior to dialysis in the binding buffer (1 mM sodium phosphate 150 mM NaCl, pH 7.4).

Workup

  1. customRecombinant purified proteins
  2. customReaction
  3. customreaction
  4. concentration1 molar ratio with respect to the protein concentration
  5. waitThe mixture was left
  6. customto react at room temperature for an hour
  7. customLabelled protein was separated from the free fluorescein on a PD-10™ column (Pharmacia)
  8. customto dialysis in the binding buffer (1 mM sodium phosphate 150 mM NaCl, pH 7.4)

Sources & evidence

ORD record
ord-718d3f64db6b42dd82e3f1b0494ba369
Source dataset
uspto-grants-2011_06
Patent number
US07960529B2

Reaction notes

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