A | B | C | D | |
Reagent stock | Stock concentration | Working concentration | Preparation notes | |
HEPES pH 7.0 / MES pH 6.0 | 0.5 M | 50 mM | ||
Bovine liver catalase | 21 mg/ml | 0.6 mg/ml | ||
DTT | 10 mM | 1 mM | Prepare fresh | |
Ascorbic acid | 40 mM | 500 µM | Prepare fresh | |
FeSO4 | 1 mM | 50 µM | Prepare 500 mM in 20 mM HCl, then dilute to 1 mM in water just before use . | |
-ketoglutarate | 20 mM | 0.5 mM | ||
Peptide | 20 mM | 100 µM | Dissolved in DMSO | |
PHD enzyme source | 20 µM | 10 µM | Prepare fresh | |
Trichloroacetic acid (TCA) | 10% | 5% | ||
Sodium phosphate pH 7.2 | 0.5 M | 50 mM | ||
2,4-DNPH | 50 mM | 25 mM | Prepare fresh. Dissolve in 0.5 M phosphoric acid, let stand for 30-60 mins, then add water to intended concentration. Filter through 0.45 µM filter to remove precipitate. | |
NaOH | 6 M | 2 M |