ArticleEuropean biophysics journal : EBJ2021
A standard operating procedure for an enzymatic activity inhibition assay.
Article in European biophysics journal : EBJ, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- Integrating In Vitro Bioactivities and In Silico Molecular Evaluation ofMolecules (Basel, Switzerland) · 2026Article
- Structural and biochemical insights into the inhibition ofRSC chemical biology · 2026Article
- Acidosis Drives Vasculogenic Mimicry in PDAC CSCs via Na+/H+ Exchanger Isoform 1 (NHE1) and Calcium Entry.Cells · 2026Article
- In silico discovery of thioglycoside analogues as donor-site inhibitors of glycosyltransferase LgtC.Scientific reports · 2026Article
- Novel prolyl endopeptidase inhibitor fromRSC advances · 2025Article
- Synthesis and biological evaluation of novel peptidomimetic inhibitors of the coronavirus 3C-like protease.European journal of medicinal chemistry · 2024Article
- Fluorine-induced polarity increases inhibitory activity of BPTI towards chymotrypsin.RSC chemical biology · 2022Article
- Community-building and promotion of technological excellence in molecular biophysics: the ARBRE-MOBIEU network.European biophysics journal : EBJ · 2021Article
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Authors and funding
4 authors.
Funding
Abstract
This Standard Operating Protocol (SOP) describes the key steps of experimental setup for an inhibition assay of enzymatic activity. The protocol begins with the design of an experiment, including the choice of a catalytic reaction, optimal conditions, fraction and concentration of the active enzyme, substrate and inhibitor concentrations and the positive and negative controls. The protocol ends with the data analysis followed by a typical example of an experiment. Despite an apparently standard procedure, the assay has a number of possible pitfalls such as low fraction of the active enzyme or errors in the analysis such as application of an improper model or incorrect determination of the inhibition constant while not recognizing the dependence on enzyme concentration. The protocol provides examples of necessary steps and controls to avoid these problems and obtain highly reliable results.
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