FLUOR DE LYS^®-Green HDAC fluorometric activity assay kit



BML-AK530-0001	96 wells	Inquire for pricing
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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. It comes in a convenient 96-well format, with all reagents necessary for fluorescent HDAC or sirtuin activity measurements and calibration of the assay. 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.

FLUOR DE LYS®-Green HDAC fluorometric activity assay kit image

Figure 1: Reaction scheme of the FLUOR DE LYS^®-Green HDAC assay kit. Deacetylation of the substrate sensitizes it to the developer, which then generates a fluorophore (lily-symbol).

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Product Details

Alternative Name:	Histone deacetylase green fluorescent assay kit

Applications:	Fluorescent detection, HTS Activity assay, Cell-based assays

Handling:	Store all components (except microtiter plates) at -70°C. Unused HeLa Nuclear Extract should be refrozen quickly, if possible snap freeze in liquid nitrogen.

Shipping:	Dry Ice

Long Term Storage:	-80°C

Contents:	Nuclear Extract from HeLa Cells (human cervical cancer cell line) (Prod. No. BML-KI140) (100µl in 0.1M potassium chloride, 20mM HEPES/NaOH, 20% (v/v) glycerol, 0.2mM EDTA, 0.5mM DTT, 0.5mM PMSF) Storage: -70°C. FLUOR DE LYS^®-Green Substrate (Prod. No. BML-KI572) (50µl 50mM in DMSO) Storage: -70°C. FLUOR DE LYS^® Developer Concentrate (20X) (Prod. No. BML-KI105) (300µl 20x stock solution; dilute in assay buffer before use) Storage: -70°C. Trichostatin A (Prod. No. BML-GR309) (100µl 0.2mM in DMSO) Storage: -70°C. FLUOR DE LYS^®-Green Standard (Prod. No. BML-KI605) (30µl 1mM in DMSO) Storage: -70°C. Note: BML-KI605 is an improved replacement of BML-KI573. NAD⁺ (Sirtuin Substrate) (Prod. No. BML-KI282) (500µl; 50mM in 50mM TRIS, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -70°C. Nicotinamide (Prod. No. BML-KI283) (500µl; 50mM in 50mM TRIS, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -70°C. HDAC Assay Buffer (Prod. No. BML-KI143) (20ml; 50mM TRIS, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -70°C. 96 Well Plates (Prod. No. BML-KI101) Storage: Room temperature White NBS Microplate (Prod. No. BML-KI571) Storage: Room temperature

Technical Info/Product Notes:	Biocompare Article: Keep an Eye on Apoptosis with Caspase Assays

Regulatory Status:	RUO - Research Use Only

Product Literature References

Rational design, synthesis and preliminary antitumor activity evaluation of a chlorambucil derivative with potent DNA/HDAC dual-targeting inhibitory activity: R. Xie, et al.; Bioorg. Med. Chem. Lett. 27, 4415 (2017), Abstract;

Synthesis and evaluation of novel dual BRD4/HDAC inhibitors: S. Amemiya, et al.; Bioorg. Med. Chem. 25, 3677 (2017), Abstract;

Various Lamin A/C mutations alter expression profile of mesenchymal stem cells in mutation specific manner: A. Malashicheva, et al.; Mol. Genet. Metab. 115, 118 (2015), Application(s): Measurement of whole histone deacetylase activity, Abstract;

Sir2 is required for Clr4 to initiate centromeric heterochromatin assembly in fission yeast: B.J. Alper, et al.; EMBO J. 32, 2321 (2013), Abstract; Full Text

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