/
/
Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase
È necessario avere un abbonamento a JoVE per visualizzare questo. Accedi o inizia la tua prova gratuita.
Capitoli
- 00:05Titolo
- 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
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.
