Total cellular HDAC activity was measured with the Fluor-de-Lys HDAC Fluorometric Cellular activity assay kit (Enzo Life Sciences, BML-AK503-0001). In brief, NRVMs plated in 96 wells were treated with HDAC inhibitor or HDAC RNAi and subsequently incubated with 200 µM Fluor-de-lys substrate (4h). The cell-permeable acetylated substrate fluoresces when deacetylated. Fluor-de-lys lysis buffer, trichostatin A, and developer solution were added 15 min before the end of the experiment, and fluorescence was measured in a microplate reader (Ex 360 nm, Em 460 nM).
Fluor de lys hdac fluorometric cellular activity assay kit
Manufactured by Enzo Life Sciences
The Fluor-de-Lys HDAC Fluorometric Cellular activity assay kit is a laboratory equipment product designed to measure the activity of histone deacetylases (HDACs) in cellular samples. The kit utilizes a fluorometric substrate to detect HDAC activity within a cellular environment.
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2 protocols using fluor de lys hdac fluorometric cellular activity assay kit
1
HDAC Activity Measurement in NRVMs
2
Measuring HDAC Activity in Cells
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Cells cultured with or without bacteria were treated with antibiotics as mentioned before, followed by in situ HDAC activity measurement with a FLUOR DE LYS HDAC Fluorometric Cellular Activity Assay Kit (Enzo Life Sciences).
In addition, nuclear proteins were extracted from these cells and subjected to HDAC activity evaluation with the same method. The conditioned supernatants derived from cells with indicated treatment were added into assay buffer during the incubation of recombinant HDAC1/HDAC2 (Proteintech) with assay substrates to investigate their effects on the activities of HDAC1/HDAC2. 1 μmol/L trichostatin A (TSA) supplied within the kit was used as a positive control for HDAC activity inhibition.
In addition, nuclear proteins were extracted from these cells and subjected to HDAC activity evaluation with the same method. The conditioned supernatants derived from cells with indicated treatment were added into assay buffer during the incubation of recombinant HDAC1/HDAC2 (Proteintech) with assay substrates to investigate their effects on the activities of HDAC1/HDAC2. 1 μmol/L trichostatin A (TSA) supplied within the kit was used as a positive control for HDAC activity inhibition.
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