In vitro peptidoglycan degradation assay using fluorescein-5-isothiocyanate-labelled sacculi
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We describe a method using fluorescein-5-isothiocyanate (FITC)-labelled peptidoglycan sacculi to characterize peptidoglycan hydrolases in vitro . In this protocol, soluble hydrolytic products released by peptidoglycan hydrolases are separated from the insoluble FITC-sacculi, quantified and set in relation to the overall available substrate. This enables the study of hydrolase activity in end-point or time-course experiments.
