In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones

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11:36 min

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July 25th, 2019

DOI :

10.3791/59385-v

July 25th, 2019

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Louis Masclef*¹, Uriarte Maxime*¹, Oumaima Ahmed¹, Nadine Sen Nkwe¹, Haithem Barbour¹, Nicholas V.G. Iannantuono¹, Amina Boubekeur¹, Salima Daou¹^,², El Bachir Affar¹^,²

¹Maisonneuve-Rosemont Hospital Research Center and Department of Medicine, University of Montréal, ²Lunenfeld-Tanenbaum Research Institute, Sinai Health System

Podsumowanie

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Small Molecule Inhibitors

High Throughput Screening

Rozdziały w tym wideo

0:04

Title

1:01

GSH-agarose Affinity Purification of the GST-RING1B(1-159)-BMI1 (1-109) E3 Ubiquitin Ligase Complex

2:16

Purification of the BAP1/DEUBAD (ASXL2) Deubiquitinase Complex

4:58

Purification of the Nucleosomes from HEK293T Cells

7:38

Nucleosome Ubiquitination Assay Using BMI1/RING1B E3 Ubiquitin Ligase Complex and In Vitro Nucleosomal H2A DUB Assay Using BAP1/DEUBAD

9:05

Results: Purification, Ubiquitination, and Deubiquitination

10:40

Conclusion

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