|Background||Inhibition of histone deacetylase (HDAC) has been implicated to modulate transcription and induce apoptosis or differentiation in cancer cells. However, screening compounds for HDAC inhibition has been difficult due to the lack of convenient tools for analyzing HDAC activity. The Fluorometric HDAC Activity Assay Kit provides a fast and fluorescence-based method that eliminates radioactivity, extractions, or chromatography, as used in traditional assays. The new procedure requires only two easy steps, both performed on the same microtiter plate. First, the HDAC substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (e.g., HeLa nuclear extract). Deacetylation of the substrate sensitizes the substrate, so that further treatment with the Lysine Developer produces a fluorophore. The fluorophore can be easily analyzed using a fluorescence plate reader or a fluorometer. The assay is well suited for high throughput screening applications. HDAC inhibitors and antibodies are also available separately. |
‧ Detection method- Fluorometer (Ex. = 350-380 nm and Em. = 440-460 nm)
‧ Sample type- Cell and Tissue lysates, culture media, urine, plasma and serum, as well as many other biological fluids
‧ Species reactivity- Mammalian
‧ Kit size- 100 assays
‧ Applications- Well suited for high throughput applications. HDAC inhibitors and substrates and related antibodies are also available separately.
Features and Benefits
‧ Simple two-step procedure; takes around than 1 hour
‧ Fast and convenient
‧ The assay method eliminates radioactivity/extractions/ and/or chromatography as used in the traditional assays.
HDAC Substrate [Boc-Lys(Ac)-AMC, 4 mM]
10X HDAC Assay Buffer
HDAC Inhibitor (Trichostatin A, 1 mM)
HeLa Nuclear Extract (5 mg/ml)
Deacetylated Standard [Boc-Lys-AMC, 4 mM]