|Background||Inhibition of histone deacetylases (HDACs) has been implicated to modulate transcription and to induce apoptosis or differentiation in cancer cells. However, screening HDAC inhibitory compounds has proven to be difficult over the past due to the lack of convenient tools for analyzing HDAC activity. The new Colorimetric HDAC Activity Assay Kit provides a fast and convenient colorimetric method that eliminates radioactivity, extractions, or chromatography, as used in the traditional assays. The new method requires only two easy steps, both performed on the same microtiter plate. First, the HDAC colorimetric substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (e.g., HeLa nuclear extract or your own samples). Deacetylation of the substrate sensitizes the substrate, so that, in the second step, treatment with the Lysine Developer produces a chromophore. The chromophore can be easily analyzed using an ELISA plate reader or spectrophotometer. The assay is well suited for high throughput screening applications. HDAC inhibitors and antibodies are also available separately. |
‧ Detection method- Colorimeter (400 or 405 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)-pNA, 10 mM]
10X HDAC Assay Buffer
HDAC Inhibitor (Trichostatin A, 1 mM)
HeLa Nuclear Extract (5 mg/ml)
Deacetylated Standard (Boc-Lys-pNA, 10 mM)