NanoPhotometer
®
N120/
NP80/N60/N50/C40
User Manual Version 4.3.1
94
K
INETICS
M
ETHOD
O
VERVIEW
The Kinetics application is useful for: measuring initial rates of enzyme-catalyzed reactions,
performing progress-curve analysis for complete reactions, calculating basic Michaelis-
Menten parameters for single-substrate reactions, and measuring enzyme inhibition. Simple
kinetics studies, where the change in absorbance is followed as a function of time at a fixed
wavelength, can be readily performed with the NanoPhotometer
®
. The rate of a chemical
reaction can be measured using spectrophotometric methods by studying the change in
absorbance at a fixed wavelength as a function of time. These changes in absorbance
reflect corresponding changes in the concentration of reactants or products as the reaction
progresses. The rate of many chemical reactions can be markedly accelerated by the
presence of catalysts, which remain chemically intact during the reaction. Catalysts in the
case of biochemical reactions are generally represented by enzymes, which are specialized
protein catalysts. However, a few examples of special reactions catalyzed by RNA
molecules also exist. Studying the kinetics of a reaction can reveal important details of the
catalytic mechanism involved in terms of sequence steps, transition state of reactants or
nature of enzyme inhibitors.
M
EASUREMENT
P
ROTOCOL
Note
: If a kinetic is started from a control device via WiFi connection (tablet or smartphone)
set the auto lock of the tablet or smartphone to never. Otherwise the kinetics will be
interrupted when the smartphone or tablet is locked, because of losing the WiFi connection.
1. Select the Kinetics icon on the home screen
2.
To change between NanoVolume and cuvette application, use the
Change to Cuvette/NanoVolume button below the parameter area
(NP80/C40 only).
3.
For Cuvette Application:
Select the path length depending on the used cuvette.
Options are: 0.5 mm, 1 mm, 2 mm, 5 mm and 10 mm
If it is desired to heat the sample to 37°C use the toggle switch to turn on cell holder heater.
When the cuvette holder has reached 37°C the toggle color changes to green.