HDAC Activity Assay with Inositol Phosphates

HDAC activity assays carried out as described in (10 (link)). To perform the titration, 62 nM protein was incubated with varying concentrations of inositol-1,4,5,6-tetraphosphate and inositol-1,4,5-triphosphate for 30 min at 37°C then the HDAC activity was measured using the HDAC Assay Kit (Active Motif) and read using a Victor X5 plate reader (PerkinElmer). Experiments were performed in triplicate and data were analyzed using GraphPad Prism (version 4.0, GraphPad Software, Inc.).

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Itoh T., Fairall L., Muskett F.W., Milano C.P., Watson P.J., Arnaudo N., Saleh A., Millard C.J., El-Mezgueldi M., Martino F, & Schwabe J.W. (2015). Structural and functional characterization of a cell cycle associated HDAC1/2 complex reveals the structural basis for complex assembly and nucleosome targeting. Nucleic Acids Research, 43(4), 2033-2044.

Publication 2015

HDAC Assay Kit Victor X5 plate reader

Assay Inositol Prism Protein Titration Triphosphate

Corresponding Organization :

Other organizations : University of Leicester, MRC Laboratory of Molecular Biology

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Variable analysis

independent variables

Varying concentrations of inositol-1,4,5,6-tetraphosphate
Varying concentrations of inositol-1,4,5-triphosphate

dependent variables

HDAC activity

control variables

62 nM protein
Incubation time of 30 min
Incubation temperature of 37°C

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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