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Assay-kits

SensoLyte® 520 HDAC Activity Assay Kit Fluorimetric - 1 kit

$582.00
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  • Cat.Number : AS-72084
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Histone deacetylase (HDAC) enzymes modulate gene expression through the deacetylation of lysine residues on histone proteins and act as transcriptional repressors of genes. Based on their role in cell cycling, apoptosis and differentiation, HDACs have been chosen as therapeutic targets for the treatment of cancer and neurodegenerative diseases.
The SensoLyte® 520 HDAC Activity Assay Kit provides a convenient, two-step homogeneous procedure for measuring HDAC activity using a fluorogenic substrate. In the first step, an acetylated substrate is incubated with HDAC containing samples. Deacetylation of substrate sensitizes it, so that, in the second step mixing with the HDAC developer generates a fluorophore, which can then be detected at Ex/Em= 490 nm/520 nm. The substrate included in the kit is cell-permeable, and the assay can be used to measure HDAC activity directly in cell culture in a 96-well plate without a time-consuming cell extraction step. This kit also can be used for high throughput screening of HDAC inhibitors using extracts or purified enzymes. The long wavelength fluorescence of HDAC 520 substrate is less interfered by the autofluorescence of cell components and test compounds.

Specifications

Packaging
Kits components
  • Component A: HDAC 520 substrate, Ex/Em=490 nm/520 nm: 25 mM in DMSO, 40 µL Component B: Deacetylated 520 standard, Ex/Em=490 nm/520 nm: 25 mM in DMSO, 10 µL Component C: HeLa Nuclear Extract: ~ 4 mg protein/mL, 15 µL Component D: Assay Buffer: 20 mL Component E: Trichostatin A (Inhibitor of HDAC): 300 µM in DMSO, 100 µL Component F: HDAC Developer (10X): 0.5 mL Component G: Cell Lysis Buffer: 20 mL
Chemistry
Sequence one letter code
  • Ac-RGK(Ac)-AMC
Sequence three letter code
  • Ac-Arg-Gly-Lys(Ac)-AMC
CAS registry number
  • 660846-97-9
UniProt number
  • Q13547
Properties
Absorbance (nm)
  • 490
Emission (nm)
  • 520
Storage & stability
Storage Conditions
  • Store component C at -80°C. Store all other components at -20°C. Components D and G can be stored at room temperature for convenience. Protect components A and B from light and moisture.
Activity
Application
Biomarker Target
Detection Method
Detection Limit
  • 0.78 ug/ml
Research Area
Sub-category Research Area
Usage
  • Research use

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Citations

Histone Deacetylase Inhibitors Prolong Cardiac Repolarization through Transcriptional Mechanisms.

Toxicol Sci. . 2016 Jun 02 ; 153(1) 39 | DOI : 10.1093/toxsci/kfw104

  • S. Spence

Transforming growth factor β1 inhibits bone morphogenic protein (BMP)-2 and BMP-7 signaling via upregulation of Ski-related novel protein N (SnoN): possible mechanism for the failure of BMP therapy?

BMC Medicine  . 2012 Sep 07 ; 6797(38) 2 | DOI : https://doi.org/10.1186/1741-7015-10-101

  • S. Ehnert

A novel isoform of human LZIP negatively regulates the transactivation of the glucocorticoid receptor

Mol Endocrinol. . 2009 Sep 24 ; 23(11) 1746 | DOI : 10.1210/me.2009-0009

  • H. Kang

A new synthetic HDAC inhibitor, MHY218, induces apoptosis or autophagy-related cell death in tamoxifen-resistant MCF-7 breast cancer cells

Invest New Drugs . 2011 Oct 08 ; 30(5) 1887 | DOI : 10.1007/s10637-011-9752-z

  • J.H. Park

Anticancer effects of the HDAC inhibitor, 3β,6β‑dihydroxyurs‑12‑en‑27‑oic acid, in MCF‑7 breast cancer cells via the inhibition of Akt/mTOR pathways.

Oncol. Rep. . 2023 Feb 01 ; 49(2) 43 | DOI : https://doi.org/10.3892/or.2023.8480

  • JS. Lim
  • et al