Features and Benefits

• Simple two-step sensitive and reliable assay
• All steps performed in the same cell culture plate
• Utilizes fluorometric method
• Sample type: cultured, adherent, and suspension cells
• Suitable for individual tests or high throughput assays and kinetic studies
• Convenient 96-well microplate format
• Suitable for screening HDAC inhibitors or activators
• Suitable for studying growth factors or other regulators that influence HDAC activity

Application

In Situ Histone Deacetylase (HDAC) Activity Fluorometric Assay Kit has been used to determine HDAC activity.

Biochem/physiol Actions

Site specific histone acetylation and deacetylation have been shown to activate or repress eukaryotic gene transcription, respectively, and as a consequence, histone deacetylases (HDACs) play a crucial role in organism development and disease. Mutation or abnormal expression of HDAC is observed in cancer.

General description

Histone deacetylases (HDACs) are a large family of enzymes that remove acetyl groups from histone proteins. HDACs are localized in both the cytosol and nucleus and some shuttle between the two locations.
Sigma′s InSitu HDAC Activity Fluorometric Assay Kit provides a direct, fast, fluorescence-based method to measure HDAC activity in cultured cells. The procedure requires just two steps, both performed in the original 96-well cell culture plate. First, the cell culture medium is replaced with a cell permeable HDAC Substrate, containing an acetylated lysine side chain. During the subsequent incubation, HDAC Substrate enters the cells and is deacetylated by intracellular HDAC. In the second step, Developer is added to lyse the cells and cleave the deacetylated HDAC Substrate to release a fluorophore. The fluorescence generated can be quantified at Ex/Em = 368/442 nm. The assay is well suited for either individual or high throughput screening.