In vitro peptidoglycan degradation assay using FITC labelled sacculi
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We describe a method using fluorescein-5-Isothiocyanate (FITC) labeled 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.
