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Journal / Biochemistry / Single-Step Enrichment of a TAP-Tagged Histone…
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Biochemistry

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JoVE Journal Biochemistry
Single-Step Enrichment of a TAP-Tagged Histone Deacetylase of the Filamentous Fungus Aspergillus nidulans for Enzymatic Activity Assay
 

Single-Step Enrichment of a TAP-Tagged Histone Deacetylase of the Filamentous Fungus Aspergillus nidulans for Enzymatic Activity Assay

Article DOI: 10.3791/59527-v • 09:07 min • May 1st, 2019
May 1st, 2019 •
1, 1, 2,3, 1, 1
1Division of Molecular Biology, Biocenter, Medical University of Innsbruck, 2Biology Department, Maynooth University, 3Maynooth University Human Health Research Institute

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Class 1 histone deacetylases (HDACs) like RpdA have gained importance as potential targets to treat fungal infections. Here we present a protocol for the specific enrichment of TAP-tagged RpdA combined with an HDAC activity assay that allows in vitro efficacy testing of histone deacetylase inhibitors.

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TAP-tagged Histone Deacetylase Enzymatic Activity Assay Filamentous Fungus Aspergillus Nidulans Fungal Infections Purified Enzyme Activities Rapid Separation Single Step Enrichment Native Complexes Activity Determination Fungal-specific Deactylase Inhibitors Extraction And Purification TAP-tagged Fungal Proteins Conidia Preparation CSS Flask Shaking Conidia Scraping Cell Strainers Centrifuge Tube Dilution Mycelia Preparation
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