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

11:01 min

November 23, 2016

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Chapters

  • 00:05Title
  • 01:11Preparation of 2-pyridyl Undec-10-ynyl Carbonate and Undec-10-ynyl 2-oxopyridine 1-carboxylate
  • 02:44Preparation of Undec-10-ynyl-N-[(3S)-2-oxoazetidin-3-yl]carbamate
  • 04:17Click Chemistry with Azide-PEG3-Biotin
  • 05:14Removal of Excess Click Chemistry Reagents
  • 06:29Streptavidin Enrichment
  • 07:57Click Chemistry with Azide-PEG3-Fluor 545 on Tissue Slices
  • 08:59Results: Synthesis and Applications of NAAA Activity Based Probe, ARN14686
  • 10:21Conclusion

Summary

Automatic Translation

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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