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Biochemistry

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

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

Article DOI: 10.3791/57798-v • 10:34 min • October 15th, 2018
October 15th, 2018 •
*1, *1, *1, 1, 1
1Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri-Kansas City
* These authors contributed equally

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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.

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Keywords: TET2 5-Methylcytosine Dioxygenase Protein Purification LC-MS/MS Bacterial Transformation Protein Expression IPTG Induction E. Coli BL21 DE3 Epigenetics Demethylation
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