JoVE Journal

Biochemistry

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Nanyang Technological University

Efficient Purification and LC-MS/MS-based Assay Development for Ten-Eleven Translocation-2 5-Methylcytosine Dioxygenase

Here, we present a protocol for an efficient single step purification of the active untagged human ten-eleven translocation-2 (TET2) 5-methylcytosine dioxygenase using ion-exchange chromatography and its assay using a liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based approach.

Chapters in this video

0:04

Title

0:45

Cloning and Purification of Untagged Human TET2 Dioxygenase

5:30

5mC Oxidation by TET2 Dioxygenase

7:02

Quantitative Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)-based Assay Development

8:31

Results: Investigating the Role of the TET2 Enzyme in Normal Development and Disease

9:26

Conclusion

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