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Promega Corporation

HDAC-Glo™ I/II Assay and Screening System Technical Manual

Literature # TM335

The HDAC-Glo™ I/II Assay and Screening System are single-reagent-addition, homogeneous, luminescent assays that measure the relative activity of histone deacetylase (HDAC) class I and II enzymes from cells, extracts or purified enzyme sources. This assay is broadly useful for class I and II enzymes, but sensitivity and performance will vary with catalytic efficiency of particular class I and II isoenzymes. The assay uses an acetylated, live-cell-permeant, luminogenic peptide substrate that can be deacetylated by HDAC activities. Deacetylation of the peptide substrate is measured using a coupled enzymatic system in which a protease in the Developer Reagent cleaves the peptide from aminoluciferin, which is quantified in a reaction using Ultra-Glo™ Recombinant Luciferase. The HDAC-mediated luminescent signal is persistent and proportional to deacetlyase activity, allowing batch processing of multiwell plates.

Printed in USA. Revised 9/11.

 

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