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In vitro peptidoglycan degradation assay using fluorescein-5-isothiocyanate-labelled sacculi

Lookup NU author(s): Dr Adam HartORCiD, Manuel BanzhafORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

© 2026 The Authors.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.

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Author(s): Alodaini D, Hart A, Moynihan PJ, Banzhaf M

Publication type: Article

Publication status: Published

Journal: Access Microbiology

Year: 2026

Volume: 8

Issue: 7

Online publication date: 15/07/2026

Acceptance date: 28/05/2026

Date deposited: 24/08/2026

ISSN (electronic): 2516-8290

Publisher: Microbiology Society

URL: https://doi.org/10.1099/acmi.0.001178.v3

DOI: 10.1099/acmi.0.001178.v3

Data Access Statement: All data associated with this work are reported within the article.

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