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Biochemistry

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JoVE Journal Biochemistry
Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase
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Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase

Article DOI: 10.3791/54652-v • 11:01 min • November 23rd, 2016
November 23rd, 2016 •
1, 1, 1, 1, 1, 1, 1, 1,2
1Drug Discovery and Development, Istituto Italiano di Tecnologia, 2Departments of Anatomy and Neurobiology, Pharmacology, and Biological Chemistry, University of California, Irvine School of Medicine

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Here, we describe the preparation and use of an activity-based probe (ARN14686, undec-10-ynyl-N-[(3S)-2-oxoazetidin-3-yl]carbamate) that allows for the detection and quantification of the active form of the proinflammatory enzyme N-acylethanolamine acid amidase (NAAA), both in vitro and ex vivo.

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Activity-based Probe N-acylethanolamine Acid Amidase NAAA Synthesis Detection Enzyme Cell Extracts Tissue Sections In Vivo Function Physiological Conditions Pathological Conditions Inflammatory Diseases Neurodegenerative Diseases Advantages Reliable Information Active Form Of The Enzyme Fluorescence Microscopy Synthesis Procedure
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