Reaktion #490366

ord-1fe8ee8a588749069a0fba50815696f9

Lösungsmittel

Reaktionsbedingungen

Detaillierte Bedingungen
See reaction.notes.procedure_details.

Aufarbeitung

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

Vorschrift

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.

Quelle

DOI: 10.6084/m9.figshare.5104873.v1Patent: US08748380B2uspto-grants-2014_06