A high yield microscale enzymatic synthesis and purification Of 14C-Labeled Nicotinamide Adenine Dinucleotide Phosphate (NADP+)
✍ Scribed by Andre Ronneberg; Gordon Metz; Richard Weld; Peter Roffey; Chris Craney
- Publisher
- John Wiley and Sons
- Year
- 1992
- Tongue
- French
- Weight
- 214 KB
- Volume
- 31
- Category
- Article
- ISSN
- 0022-2135
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✦ Synopsis
Abstract
Uniformly labeled (U) ^14^C nicotinamide adenine dinucleotide phosphate (NADP^+^) was synthesized by phosphorylating [U‐^14^C]nicotinamide adenine dinucleotide (NAD^+^) in the presence of immobilized NAD^+^ kinase. The 15 μCi (600 μL) synthesis consistently achieved yields between 80% and radiochemical purities greater than 95%. The [U‐^14^C]NADP^+^ was purified by high performance anion‐exchange chromatography using a gradient elution of ammonium bicarbonate. This procedure may be applicable to the synthesis of other charged, UV‐absorbing products of enzyme‐catalyzed reactions.