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JoVE Journal Biochemistry
Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening
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Journal (Biochemistry)
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Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening

Article DOI: 10.3791/61146-v • 07:37 min • May 13th, 2020
May 13th, 2020 •
1, 1, 1
1Institut Pasteur, Unité Biologie et génétique de la paroi bactérienne, Paris, France ; CNRS, UMR 2001 « Microbiologie intégrative et Moléculaire », Paris, France ; INSERM, Équipe Avenir, Paris, France

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Summary
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Presented here is a sensitive fluorescence assay to monitor apolipoprotein N-acyltransferase activity using diacylglyceryl peptide and alkyne-phospholipids as substrates with click-chemistry.

Tags

Click-Chemistry Fluorometric Assay Apolipoprotein N-acyltransferase Enzyme Characterization High-Throughput Screening HDS Format Inhibitors Protocol Sensitive Assay Screening Purposes Potential Novel Antibiotics Mixing Vortexing Sonication Nonhomogeneous Lipid Detergent Containing Suspensions Reagent Mixture Alkyne POPE FSL-1-biotin LNT Reaction Buffer Triplicate Reagent Mixtures Sonication Ultrasonic Water Bath Incubation Active LNT Inactive LNT Concentration Thermal Mixer Heated Lid Stock Solution
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