Products from BPS Bioscience require a minimum order value above 400€
Applications: Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Background: HDACs regulate cellular processes by catalyzing the hydrolysis of an acetyl group fromacetyllysines in modified proteins. In the HDAC assay, fluorescent-dye molecules areattached to a peptide containing acetyllysine. Attachment to the peptide quenches thefluorescence of the dye. After treatment of the peptide with an HDAC, the reaction ismixed with a development solution that is specific for nonacetylated lysines. If the acetylgroup has been removed from the lysine by the HDAC, this solution will release the dyeallowing for fluorescence. Fluorescence is therefore directly related to HDAC activity.
Contraindications: DMSO >1%, strong acids or bases, ionic detergents, high salt
Description: The Fluorogenic HDAC Assay Kit is a complete assay systemdesigned to measure histone deacetylase (HDAC) class 1 activity for screeningand profiling applications. The kit comes in a convenient 96-well format, with all thereagents necessary for 100 fluorescent HDAC activity measurements. Inaddition, the kit includes purified HDAC2 enzyme and a potent HDAC inhibitor,Trichostatin A, for use as a positive and negative control. The Fluorogenic HDACAssay Kit is based on a unique fluorogenic substrate and developer combination.This assay method eliminates dealing with the radioactivity, extraction, andchromatography aspects of traditional assays. Using this kit, only two simplesteps on a microtiter plate are needed to analyze the HDAC activity level. First,the HDAC fluorometric substrate, containing an acetylated lysine side chain, isincubated with purified HDAC enzyme. The deacetylation sensitizes thesubstrate so subsequent treatment with the Lysine Developer produces afluorophore that can then be measured using a fluorescence reader at 485 nm(excitation)/528 nm (emission).Our other HDAC kit (#50061) contains a substrate that is excited at wavelengths 350-380 nm and fluoresces at wavelengths 440-460 nm. However, the substrate in this kit fluoresces at longer (green) wavelengths: 485 nm(excitation)/528 nm (emission). It is most useful when the sample or inhibitor fluoresces at wavelengths that overlap those of our other HDAC substrate
Storage Stability: 12 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. A. Ito et al., 2001, EMBO J. 20 1331.
2. N.A. Barlev et al., 2001, Mol. Cell 8 1243.
3. A. Ito et al., 2002, EMBO J. 21 6236.Application References:>Mitogen-Activated Protein Kinase Phosphatase 3 (MKP-3)-Deficient Mice Are Resistant to Diet-Induced Obesity (2014)Investigating the selectivity of metalloenzyme inhibitors (2013)
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