Colorimetric Assay for Apyrase Activity

The apyrase activity was measured using a colorimetric microassay based on the Fiske and Subbarow method [55 (link)] with slight modifications [56 (link)] on the Infinite M 200 Fluorometer (Tecan, Männedorf, Switzerland) at 665 nm. The concentration of the Pi was calculated from a standard curve with potassium dihydrogen phosphate. One unit of enzyme activity was defined as the amount of enzyme that releases one micromole of orthophosphate per minute from the nucleotide substrate at the specified assay conditions.

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Polanska N., Ishemgulova A., Volfova V., Flegontov P., Votypka J., Yurchenko V, & Volf P. (2020). Sergentomyia schwetzi: Salivary gland transcriptome, proteome and enzymatic activities in two lineages adapted to different blood sources. PLoS ONE, 15(3), e0230537.

Publication 2020

Infinite M 200 Fluorometer

Apyrase Assay Colorimetric Enzyme Enzyme activity M 200 Nucleotide Orthophosphate Potassium dihydrogen phosphate Releases

Corresponding Organization : Charles University

Other organizations : University of Ostrava, Czech Academy of Sciences, Biology Centre, Institute of Parasitology, Sechenov University

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Variable analysis

independent variables

Concentration of the Pi (potassium dihydrogen phosphate)

dependent variables

Apyrase activity

control variables

Assay conditions (temperature, pH, etc.)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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