CycLex HDACs Deacetylase Fluorometric Assay Kit Ver.2

Histone deacetylase (HDAC) is considered to play a crucial role in regulating gene expression by changing nucleosome structure. HDAC is also thought to participate in regulation of cell cycle and differentiation, and it has been reported that the failure of this regulation leads to some types of cancer. Inhibition of HDAC activity by HDAC inhibitors such as Trichostatin A (TSA) and suberoylanilide hydroxamic acid (SAHA) induce differentiation and/or apoptosis of transformed cells in vitro and inhibit tumor growth in a mouse model. It has been reported that HDAC inhibitors are effective for the medical treatment of acute promyelocytic leukemia (APL) and various cancers. Thus, HDAC inhibitors are expected to function as new anti-tumor drugs and antibacterial reagents. It is thought that screening of histone deacetylase inhibitors is likely to be further carried out, as one way to discover additional substances with similar properties. However, the conventional method for measuring HDAC activity is very complicated and laborious. In order to measure HDAC enzyme activity, it is necessary to prepare radioactive acetylated histone as a substrate. First, cells have to be labeled metabolically with radioactivity by adding radioactive acetic acid to the culture medium. Second, radioactive acetylated histone has to be purified from the cells. Following the reaction, it is necessary to extract and separate the radioactive acetyl group, which has been released from acetylated histone, using ethyl acetate to measure the activity of the enzyme based on the radioactivity. Although a method for measuring the activity of deacetylase without the use of radioactive substances was reported in recent years, owing to the use of fluorescent-labeled acetylated lysine as a substrate, the reaction product must be separated from the intact substrate and the fluorescent intensity measured by reverse phase HPLC. As mentioned above, these measurement systems are difficult to adapt for processing many samples under a variety of conditions, because of their complicated operation. Thus a simple system for biochemical analysis as well as for inhibitor screening without the use of radioactive substances is preferred.

The CycLex Research Product CycLex HDACs Deacetylase Fluorometric Assay kit detects HDAC activity in lysates. Primarily, the CycLex Research Product CycLex HDACs Deacetylase Fluorometric Assay kit is designed for the rapid and sensitive evaluation of HDAC inhibitors using crude HDAC fraction. Additionally, any cultured primary cell, cell line, or tissue homogenate can be assayed for HDAC activity with the CycLex Research Product CycLex HDACs Deacetylase Fluorometric Assay kit if the appropriate dose of HDAC specific inhibitor e.g. Trichostatin A is used.

HDAC Assay Buffer, Fluoro-Substrate Peptide (0.2 mM), Fluoro-Deacetylated Peptide (0.2 mM), Trichostatin A (10 uM), Developer, HDACs, Stop Solution

1 year

HDACs