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| | HDAC Activity screening |
| Description: | Histone Deacetylase (HDAC) Inhibition Assay using Boc-Lys(Ac)-AMC Substrate: Inhibition of HDAC has been implicated to modulate transcription and to induce apoptosis or differentiation in cancer cells. The fluorometric assay provides a fast and fluorescence based method that eliminates radioactivity, extractions or chromatography, as used in traditional assays. The assay is based on two steps. First, the HDAC fluorometric substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (Mouse Liver Extract). Deacetylation of the substrate sensitizes the substrate; in the second step, treatment with the Trypsin, stop solution produces a fluorophore that can be easily analyzed using fluorescence plate reader. Assay was done in 96-well black microplate and total volume of the assay was 100 mL. Mouse liver enzyme (10 mg/mL) was diluted 1:6 with HDAC buffer. Enzyme cocktail was made of 10 mL of diluted enzyme and 30 mL of HDAC buffer. Forty m |
| Affinity data for this assay | |
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