NanoPhotometer
®
NP80/N60/N50/C40
User Guide Version 3.1
66
C
ALCULATIONS
Protein concentration is determined using the standard curve by correlating absorbance values
of samples with known concentration to calculate the concentration of the unknown sample. In
order to maintain accuracy and precision please ensure that the R
2
value of the standard curve
is 0.95 or greater.
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
