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Improvement and Validation of the Fluorescence‐Based Histone Deacetylase Assay Using an Internal Standard
Author(s) -
Hoffmann Katharina,
Heltweg Birgit,
Jung Manfred
Publication year - 2001
Publication title -
archiv der pharmazie
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 61
eISSN - 1521-4184
pISSN - 0365-6233
DOI - 10.1002/1521-4184(200107)334:7<248::aid-ardp248>3.0.co;2-k
Subject(s) - histone deacetylase , chemistry , histone , histone deacetylase inhibitor , acetylation , hdac10 , hdac11 , fluorescence , biochemistry , histone deacetylase 5 , microbiology and biotechnology , gene , biology , physics , quantum mechanics
The determination of the activity of histone deacetylase (HDAC) and the potency of its inhibitors has become an important goal in medicinal chemistry. This is due both to the involvement of HDAC in gene regulation and the ability of its inhibitors to modulate transcription and induce differentation and/or apoptosis in cancer cells. We have previously reported the development of a non‐isotopic assay for HDAC using a fluorescent derivative of ε‐acetyl lysine. It can replace existing methods that rely on radioactively labeled histones or oligopeptides as substrates. Here we report validation and improvement of the procedure using an internal standard for the quantitation of the fluorescent substrate by HPLC.