Product Details
Form: Crystalline powder
Molecular Weight: 663.4
Solubility: Distilled water or dilute buffer
Stability: Store at -20° C (-4° F)
Purity: >95%
Applications
NAD is used in the determination of amylase, creatine kinase, transaminases, lactate dehydrogenase, phosphohexose isomerase, ethanol, galactose, glucose, uric acid, L-lactate, triglycerides and other enzymes and metabolites.
Reagents
- Sodiumdiphophate/semicarbazide/substrate, pH 8.7: 3.33 g Na4P2O7&10 H2O, 0.84 g semicarbazide&HCl, 0.17 glycine, 1.00 ml ethanol with 80 ml distilled water. Adjust pH to 8.7 with 2 M NaOH; adjust volume to 100 ml.
- Alcohol dehydrogenase, from yeast (30 mg protein/ml): 300 U/ml.
Procedure
- Dissolve 25 mg NAD in 25 ml distilled water in a volumetric flask.
- Set spectrophotometer (equipped with strip chart recorder and temperature control) at 340 nm and 25°C.
- Into a cuvette, pipette the following:
Buffer (1) 3.00 ml
sample 0.10 ml
- Mix and read the absorbance A1
- Start the reaction by adding 0.01 ml of Alcohol dehydrogenase (2). Mix and read absorbance A2.
- Add another 0.01 ml Alcohol dehydrogenase. Mix and read absorbance A3.