This video demonstrates the dye release assay for quantifying the enzymatic activity of antimicrobial proteins. The antimicrobial protein degrades the bacteria's dye-labeled cell wall and releases the dye molecules turning the solution blue. The absorbance of the solution correlates with the enzymatic activity of the antimicrobial protein.
Protocol
1. Preparation of Bacteria NOTE: Whole-cell bacterial substrates and purified peptidoglycans are used as enzyme substrates in both the microslide diffusion assay and the dye-release assay. These substrates require preparation prior to conducting the enzyme reactions. The following protocol describes their preparation. Whole Bacterial Cell Substrate Produce bacterial cell substrate by inoculating 500 ml of nutrient broth with a 2 ml over…
Disclosures
The authors have nothing to disclose.
Materials
48-well flat-bottom microplate with low evaporation lid