A stopped-flow mixer device for a batch microcalorimeter application to NAD-NADase reaction

J Biochem Biophys Methods. 1989 Mar;18(2):113-24. doi: 10.1016/0165-022x(89)90073-0.

Abstract

A new molded polypropylene, diamond-like carbon (DLC)-coated mixing cell has been developed for use in the batch microcalorimeter. Reagent volume can be varied from 25 microliters to 100 microliters. A 10 microcalorie reaction heat can be measured to 5%. Repeat reactions can be done as often as every 10 min for a fast reaction. Reactions can be started within 1 h or less after loading. A pre-equilibrator and a temperature-controlled syringe drive unit permit solutions to be stored at 4 degrees C while being run at any temperature from -20 degrees C to 40 degrees C. The kinetics and enthalpy of reaction of NAD-NADase have been measured. delta H is about 21 kcal/mol endothermic.

MeSH terms

  • Calorimetry / instrumentation
  • Calorimetry / methods
  • Kinetics
  • Microchemistry
  • NAD / metabolism*
  • NAD+ Nucleosidase / metabolism*

Substances

  • NAD
  • NAD+ Nucleosidase