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

HRID 2358369Notes

Reaction equation

HRID 2358369 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-adad73df764f4abe8b286c4ad6b50a27
Source dataset
uspto-grants-2010_07
Patent number
US07763712B2

Reaction notes

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