PCI-34051

Histone deacetylase activity

For PCI-34051 characterization, measurements were perfomed in a reaction volume of 100 μL using 96-well assay plates in a fluorescence plate reader. For each isozyme, the HDAC protein in reaction buffer (50 mM HEPES, 100 mM KCl, 0.001% Tween-20, 5% dimethyl sulfoxide, pH 7.4, supplemented with bovine serum albumin at concentrations of 0 ~ 0.05%) was mixed with PCI-34051 at various concentrations and allowed to incubate for 15 mins. Trypsin was added to a final concentration of 50 nM, and acetyl-Gly-Ala-(N-acetyl-Lys)-amino-4-methylcoumarin was added to a final concentration of 25 ~ 100 μM to initiate the reaction. After a 30 min lag time, the fluorescence was measured over a 30 min time frame using an excitation wavelength of 335 nm and a detection wavelength of 460 nm. The increase in fluorescence with time was used as the measure of the reaction rate.

Cell lines

T-cell derived lines (Jurkat, HuT78, HSB-2 and Molt-4 cells)

Preparation method

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reaction Conditions

5 μM; 2 days

Applications

The HDAC8-selective inhibitor PCI-34051 was cytotoxic to T-cell derived lines.

References:

[1]. Balasubramanian S, Ramos J, Luo W, Sirisawad M, Verner E, Buggy JJ. A novel histone deacetylase 8 (HDAC8)-specific inhibitor PCI-34051 induces apoptosis in T-cell lymphomas. Leukemia. 2008 May;22(5):1026-34.