HDAC Activity Assay Kit (BN00597)

SKU:
BN00597
Product type:
Cellular Assay
Detection Method:
Fluorometric
€565

Description

Technical Manual MSDS

HDAC Activity Assay Kit Data

Assay Information Description
Product Name: HDAC Activity Assay
Product SKU: BN00597
Size: 100 assays
Application:
  • Well suited for high throughput applications
  • HDAC inhibitors and substrates and related antibodies are also available separately
Detection Method: Fluorometric (Ex/Em 350-380/440-460 nm)
Reactivity: Mammalian
Assay Time: < 1 hour (Simple two step procedure)
Storage: Store the kit at -80°C, protect from light
Shipping Conditions: DRY  Ice
Usage: For research use only. Not for use in humans.

HDAC Activity Assay Kit Components

Component Quantity
HDAC Substrate 500 µl
10X HDAC Assay Buffer 1 ml
Lysine Developer 1 ml
HDAC Inhibitor (Trichostatin A, 1 mM) 10 µl
HeLa Nuclear Extract (5 mg/ml) 10 µl
Deacetylated Standard (4 mM) 20 µl

Product Images

Figure: Analyses of HDAC Activity in HeLa Nuclear Extract. HeLa nuclear extract (NE) in various amounts were incubated with 5 µl HDAC fluorometric substrate. After 30 min, the reactions were stopped with 10 µl Lysine Developer. Samples were then read in a fluorescence plate reader with Ex/Em = 360/460 nm.

HDAC Activity Assay Kit Principle

Inhibition of histone deacetylase (HDAC) has been implicated to modulate transcription and induce apoptosis or differentiation in cancer cells. However, screening compounds for HDAC inhibition has been difficult due to the lack of convenient tools for analyzing HDAC activity. The Fluorometric HDAC Activity Assay Kit provides a fast and fluorescence-based method that eliminates radioactivity, extractions, or chromatography, as used in traditional assays. The new procedure requires only two easy steps, both performed on the same microtiter plate. First, the HDAC substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (e.g., HeLa nuclear extract). Deacetylation of the substrate sensitizes the substrate, so that further treatment with the Lysine Developer produces a fluorophore. The fluorophore can be easily analyzed using a fluorescence plate reader or a fluorometer. The assay is well suited for high throughput screening applications. HDAC inhibitors and antibodies are also available separately.

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