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.
Sample type:
Cell and Tissue lysates, culture media, urine, plasma and serum, as well as many other biological fluids.
Intended Use:
Measurment of HDAC in mammal samples.
Specificity:
Mammalian
Kit Components:
HDAC substrate (10mM), 1x0.5ml
10x HDAC assay buffer, 1x1ml
Lysine developer, 1x1ml
HDAC inhibitor (1mM), 1x10ul
HeLa nuclear extract (5mg/ml), 1x50ul
Deacetylated standard (10mM), 1x20ul
Storage and Stability:
Store liquid at -20 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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