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

HRID 403184Notes

Reaction equation

HRID 403184 reaction equation
[CH:1]1[C:6](N2C(=O)C=CC2=O)=[CH:5][C:4]2[C:14]([O:16][C:17]3([C:27]4[CH:28]=[CH:29][C:30]([OH:32])=[CH:31][C:26]=4[O:25][C:19]4[CH:20]=[C:21]([OH:24])[CH:22]=[CH:23][C:18]3=4)[C:3]=2[CH:2]=1)=[O:15]>CN(C)C=O>[CH:1]1[CH:6]=[CH:5][C:4]([C:14]([OH:16])=[O:15])=[C:3]([C:17]2[C:18]3[CH:23]=[CH:22][C:21]([OH:24])=[CH:20][C:19]=3[O:25][C:26]3[C:27]=2[CH:28]=[CH:29][C:30]([CH:31]=3)=[O:32])[CH:2]=1

Participants

3

Reactant 1

HCID 122038Fluorescein 5-maleimide

Product 1

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

Reagents, catalysts & solvents 1

HCID 6228DimethylformamideSolvent

Procedure

Fluorescein-5-Maleimide, Thermo Scientific (F5M) was dissolved in dimethylformamide, to give a final concentration of 25 mg/mL. This was then further diluted into 18 mls of PBS, pH adjusted to approximately pH 6.5. To this solution either 1 ml of 200 mg/mL rHA or 1 mL of 200 mg/mL K573P variant was added. This gave an approximate 20 fold final molar excess of F5M. These samples were incubated and allowed to conjugate overnight at 4° C., in the dark, to allow the maleimide groups on the F5M to react with predominantly the free sulfhydryl, present in both albumin species.

Workup

  1. customto give a final concentration of 25 mg/mL
  2. additionThis was then further diluted into 18 mls of PBS
  3. additionTo this solution either 1 ml of 200 mg/mL rHA or 1 mL of 200 mg/mL K573P variant was added
  4. customThis gave an approximate 20

Sources & evidence

ORD record
ord-1fe8ee8a588749069a0fba50815696f9
Source dataset
uspto-grants-2014_06
Patent number
US08748380B2

Reaction notes

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