Description

Histone deacetylases (HDACs) play a central role in controlling cell cycle regulation, cell differentiation, and tissue development. These proteins have crucial roles in development and physiology, especially in the heart and nervous system. They are also deeply involved in cellular proliferation, cell cycle and apoptosis. HDAC3 is primarily localized in the nucleus, but can also be found in the cytoplasm and at the plasma membrane. The HDAC3 IP & Activity Assay Kit provides an antibody-based method to specifically immunoprecipitate the HDAC3 complex from cells & tissues and to measure HDAC3 activity fluorometrically. HDA3 is immunoprecipitated from cell or nuclear extract(s) using HDAC3 antibody followed by capturing the complex on protein-A/G beads. The immunoprecipitated complex is incubated with the HDAC substrate. Only the deacetylated substrate is cleaved by the Developer to produce a fluorophore, which can be easily analyzed using a fluorescence plate reader.

Datasheet

FAQ

Does the kit detect the total activity of HDAC or the activity of HDAC 3?
The HDAC3 antibody in K344 is a high affinity polyclonal antibody that recognizes multiple epitopes on the HDAC3 protein. Hence it should only specifically pull down HDAC2. The substrate is an HDAC substrate, not a HDAC-3 specific substrate. There is always overlap in the acetylation ability of HDACs. So the point of quality control is the antibody pull-down.

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.

Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only

CITATIONS
Kazunori Inoue, Podocyte histone deacetylase activity regulates murine and human glomerular diseases. J Clin Inves, Feb 2019; 30776024.

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