of Goettingen (Germany) RUDN University, Moscow (Russian Federation) Russian Academy of Sciences (RAS), Pushchino (Russian Federation)
HELICON REMOTE REBEL T6 SERIES
TLZ enabled identifying and using linear parts on MUF time series and accounting for MUF losses in each zymogram pixel, considerably improving the accuracy as compared to the traditional method. The results indicated that the enzyme diffusion within the membrane-soil system was negligible, and more » measured zymograms were adequately reproduced solely by accounting for substrate and product diffusions and for catalytic enzyme reaction described by the Michaelis-Menten equation.
The calibrated model was used to analyze the processes governing spatial and temporal dynamics of MUF contents in the membrane.
HELICON REMOTE REBEL T6 SOFTWARE
MUF: 4-methylumbelliferone) from plant roots, enzyme activity measurements on the roots, and HYDRUS-2D & HP2 software calibration with obtained data. We assessed the new technique in a series of laboratory and modeling experiments, including quantification of the fluorescent product diffusion (e.g. In this work we introduce a new zymography technique, time-lapse zymography (TLZ) the approach that eliminates the need for assumptions of the traditional zymography and provides more realistic estimates of enzymatic activities. These assumptions are unlikely to hold in experimental settings. The traditional zymography is based on assumptions that fluorescence on membrane images is linearly increasing with time during zymography and an increase in product content is numerically equal to enzyme activity in soil below the membrane. The method consists of incubating a membrane saturated with an enzyme-specific fluorogenic substrate on the soil/root surface followed by measurements of fluorescence intensity of the product in the membrane. Membrane zymography is commonly used in rhizosphere ecology to map enzyme activities in intact soil samples and plant roots.
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