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.
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 tissue and cell samples:
Specificity:
Recognizes mammalian HDAC
Kit Components:
HDAC substrate (4mM), 1x0.5ml
10X HDAC assay buffer, 1x1ml
HDAC inhibitor (1mM), 1x10ul
HeLa Nuclear Extract (5mg/ml), 1x10ul
Deacetylated standard (4mM), 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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