Tribioscience beta-Nicotimamide Adenine Dinucleotide, NAD (TBP0095)

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

1. 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.
2. Alcohol dehydrogenase, from yeast (30 mg protein/ml): 300 U/ml.

Procedure

1. Dissolve 25 mg NAD in 25 ml distilled water in a volumetric flask.
2. Set spectrophotometer (equipped with strip chart recorder and temperature control) at 340 nm and 25°C.
3. Into a cuvette, pipette the following:

Buffer  (1)     3.00 ml

sample          0.10 ml

4. Mix and read the absorbance A1
5. Start the reaction by adding 0.01 ml of Alcohol dehydrogenase (2). Mix and read absorbance A2.
6. Add another 0.01 ml Alcohol dehydrogenase. Mix and read absorbance A3.