An improved Fluor de Lys® HDAC assay with Fluor de Lys®-Green, a new substrate offering higher sensitivity and an excitation and emission (485/530nm) that avoids quenching and fluorescent interference from compounds absorbing in the near UV and blue range.The Fluor de Lys®-Green HDAC assay is a complete kit for measuring histone deacetylase (HDAC) activity in cell or nuclear extracts, immunoprecipitates or purified enzymes
The Fluor de Lys®-Green HDAC assay is based on the Fluor de Lys®-Green substrate and Fluor de Lys® developer combination. The Fluor de Lys® system (Fluorogenic Histone deAcetylase Lysyl Substrate/Developer) is a highly sensitive and convenient alternative to radiolabeled, acetylated histones or peptide/HPLC methods for the assay of histone deacetylases. The assay procedure has two steps. First, the Fluor de Lys®-Green substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (HeLa nuclear or other extract, purified enzyme, bead-bound immunocomplex, etc.). Deacetylation of the substrate sensitizes the substrate so that, in the second step, treatment with the Fluor de Lys® developer produces a fluorophore.
It comes in a convenient 96-well format, with all reagents necessary for fluorescent HDAC or sirtuin activity measurements and calibration of the assay.
Delivery information: Nuclear Extract from HeLa Cells (human cervical cancer cell line), Fluor de Lys®-Green Substrate (50 µL 50 mM in DMSO), Fluor de Lys® Developer Concentrate (20X) (300 µL 20x stock solution; dilute in assay buffer before use), Trichostatin A (100 µL 0.2 mM in DMSO), Fluor de Lys®-Green Standard (30 µL 1 mM in DMSO), NAD+ (Sirtuin Substrate) (500 µL), Nicotinamide (500 µL), HDAC Assay Buffer (20 mL), 96 Well Plates and White NBS Microplate.
