Eosin Y as an Internal Standard for a Plate Reader-Based Quantitation of a Histone Deacetylase Substrate

• Published in: Archiv der Pharmazie (Weinheim) Vol. 335; no. 6; pp. 296 - 300
• Main Authors: Heltweg, Birgit, Jung, Manfred
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.06.2002; WILEY‐VCH Verlag; Wiley-VCH
• Subjects: Analysis; Antineoplastic Agents - pharmacology; Biological and medical sciences; Calibration; Drug Screening Assays, Antitumor; Enzyme Inhibitors - pharmacology; Eosin Y; Eosine Yellowish-(YS) - chemistry; Fluorescent Dyes - chemistry; Fluorogenic substrate; General pharmacology; Histone deacetylase; Histone Deacetylase Inhibitors; Histone Deacetylases - metabolism; Kinetics; Medical sciences; Pharmacology. Drug treatments; Reference Standards; Reproducibility of Results; Trichostatin A; Substrate; Acyltransferases; α-Aminoacid; Chemical analysis; Enzymatic activity; Enzyme; Transferases; Coumarine derivatives; Internal standard; Fluorescence; Histone acetyltransferase; Xanthene dye
• ISSN: 0365-6233; 1521-4184
• DOI: 10.1002/1521-4184(200208)335:6<296::AID-ARDP296>3.0.CO;2-6

Ongoing interest in histone deacetylase (HDAC) inhibitors as potential anticancer drugs and mechanistic tools for the study of gene regulation is driving the improvement of assay techniques for the determination of HDAC activity. We previously reported the first non‐isotopic substrate for HDAC. A plate reader‐based determination of the substrate conversion utilized a boraindacene as an internal standard which is no longer commercially available. We report here that Eosin Y is a suitable replacement for that purpose, leading to a validated HDAC assay with increased throughput.