JoVE Journal

Biochemistry

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Optimized Incorporation of Alkynyl Fatty Acid Analogs for the Detection of Fatty Acylated Proteins using Click Chemistry

Transcript

The study of fatty acylation has important implications in cellular protein interactions and diseases. Presented here is a modified protocol to improve click chemistry detection of fatty acylated proteins, which can be applied in various cell types and combined with other assays, including pulse-chase and mass spectrometry.

Chapters in this video

0:04

Introduction

0:44

Cell Culture

1:25

Preparation and Saponification of Fatty Acid Analogs

3:35

Click Reaction on Cell Lysates

4:12

Click Reaction on Immunoprecipitated Proteins

5:13

Results: Detection of Fatty Acylated Proteins Using Click Chemistry

6:57

Conclusion

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