This assay conveniently detects HDAC
activity in cell lysates and solutions, or screens for HDAC inhibitor in cell
extracts and purified enzymes. It uses a non-peptide HDAC substrate that, in
comparison to peptide-based HDAC substrate, demonstrates more resistance to
protease hydrolysis as well as higher sensitivity. This non-peptide substrate
is cleaved into a highly fluorescent green product with long wavelength
emission, allowing for less interference with cell components and other
compounds. Signals increase with greater HDAC activity, and can be read with
a fluorescence microplate reader with excitation at 490 nm and emission at
525 nm. |