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FLUOR DE LYS®-Green HDAC fluorometric activity assay kit

 
BML-AK530-0001 96 wells 459.00 USD
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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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FLUOR DE LYS®-Green HDAC fluorometric activity assay kit image

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:Shipped on 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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