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InSitu HDAC Activity Fluorometric Assay Kit

Two Easy Steps, HTS
Catalog #: K339

In stock

$345.00

Product Details

Cat # +Size K339-100
Size 100 assays
Detection Method Fluorescence (Ex/Em = 368/442 nm)
Applications Measurement of HDAC activity in cells in 96-well format
Screening HDAC inhibitors or activators
Studying growth factors or other regulators that influence HDAC activity
Features & Benefits • Simple procedure; just requires two easy steps
• Direct, Fast, Fluorescence based
• Kit contains all necessary reagents for measuring InSitu HDAC activity in adherent or suspension cells
Kit Components • HDAC Assay Buffer
• HDAC Substrate
• Developer
• HDAC Inhibitor (Trichostatin A [TSA], 1 mM)
• Positive Control (Jurkat Cell Lysate)
• Standard (Deacetylated Substrate, 4 mM)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Histone acetylases (HAT’s) and Histone deacetylases (HDAC’s) are associated with regulation of gene expression. In general, increased levels of histone acetylation are associated with increased transcriptional activity, whereas decreased levels of acetylation are associated with repression of gene expression. HDAC’s are localized in both the cytosol and nucleus and some shuttle between the nucleus and cytosol. Increased HDAC expression has been observed in various cancers. BioVision’s InSitu HDAC Activity Fluorometric Assay Kit provides a direct, fast, fluorescence-based method to measure the InSitu HDAC activity. The procedure requires two easy steps, all performed in the same cell culture plate. First, the cell permeable HDAC Substrate, which comprises an acetylated lysine side chain, is incubated with cells grown in a 96-well plate. Inside the cells, HDAC deacetylates the substrate. The second step involves lysing the cells and treating with the Developer that produces a fluorophore from the Deacetylated HDAC Substrate. The generated fluorescence can be quantified at Ex/Em = 368/442 nm. The assay is well suited for high throughput screening applications.


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What wavelength is suitable for this assay?
The fluorescence signal for this assay must be read at Ex/Em = 368/442 nm. Since all fluorometers have a window of wavelength within which the sensitivity is comparable, a +/- 20 nm difference from the recommended numbers would be fine.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
 BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
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