Fluor de Lys® Substrate is a highly sensitive and convenient alternative to radiolabeled, acetylated histones or peptide/HPLC methods for the assay of histone deacetylases. The Fluor de Lys
® Substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (nuclear or cellular extract, purified enzyme, bead-bound immunocomplex, etc.). Deacetylation of the substrate sensitizes the substrate so that, in a second step, treatment with the Fluor de Lys
® Developer produces a fluorophore.
Histone deacetylases from all known classes have been successfully assayed with the
Fluor de Lys® Substrate. These include human HDACs 1, 2, 3 and 8 (class I), human HDACs 4 and 9 (class II) and the sirtuins yeast Sir2, human SIRT1 and human SIRT2 (class III; reaction must include the sirtuin substrate NAD
+). The substrate is cell-permeable and serves as the basis for the HDAC Cellular Activity Assay Kit (Prod. No. BML-AK503).
NOTE: Must be used in conjunction with Fluor de Lys
® Developer (Prod. No. BML-KI105). Sufficient for 100-1000 assays of HDACs or sirtuins (50 µl reaction volume, 50-500 µM (substrate).
Product Specification
General Literature References
A microplate reader-based nonisotopic histone deacetylase activity assay: B. Heltweg and M. Jung; Anal. Biochem.
302, 175 (2002),
Abstract;
A molecular mechanism of action of theophylline: Induction of histone deacetylase activity to decrease inflammatory gene expression: K. Ito et al.; PNAS
99, 8921 (2002),
Abstract;
Inhibition of silencing and accelerated aging by nicotinamide, a putative negative regulator of yeast sir2 and human SIRT1: K.J. Bitterman et al.; J. Biol. Chem.
277, 45099 (2002),
Abstract;
Proliferating cell nuclear antigen associates with histone deacetylase activity, integrating DNA replication and chromatin modification: S. Milutinovic et al.; J. Biol. Chem.
277, 20974 (2002),
Abstract;
Cloning and characterization of a histone deacetylase, HDAC9: X. Zhou et al.; PNAS
98, 10572 (2001),
Abstract;
BIOMOL Research Laboratories, Inc.; Unpublished results (0),