Description
Key facts
Detection method:Colorimetric,
Sample types:Nuclear Extracts, Cell Lysate,
Assay type:Quantitative,
Assay Platform:Microplate (12 x 8 well strips)
Product details:
HDAC Activity Assay Kit (Colorimetric) ab1432 provides a fast and convenient colorimetric method that eliminates radioactivity, extractions, or chromatography, as used in the traditional assays. The new method requires only two easy steps, both performed on the same microtiter plate. First, the HDAC colorimetric substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (e.g., HeLa nuclear extract or your own samples). Deacetylation of the substrate sensitizes the substrate, so that, in the second step, treatment with the Lysine Developer produces a chromophore. The chromophore can be easily analyzed using an ELISA plate reader or spectrophotometer. The assay is well suited for high throughput screening applications.
Inhibition of histone deacetylases (HDACs) has been implicated in the modulation of transcription and the induction of apoptosis or differentiation in cancer cells. However, screening HDAC inhibitory compounds has proven to be difficult over the past due to the lack of convenient tools for analyzing HDAC activity.
Other Notes
This product was previously called K331 HDAC Activity Colorimetric Assay Kit. Biovision was acquired by Abcam in 2021.
Read the entire protocol (attached to the datasheet) before beginning the procedure.
The HeLa extract should be refrozen immediately at -20 or -70°C after each use to avoid loss of activity. The Lysine Developer should be refrozen immediately at -20 or -70°C after each use or aliquotted for future use.
This kit contains enough reagents for about 100 tests. If positive and negative controls are included, the kit provides sufficient reagents for 5 positive control assays with the HeLa Nuclear Extract and 5 Negative Control assays with the HDAC Inhibitor, Trichostatin A.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Dry Ice, Appropriate short-term storage conditions--80°C, Appropriate long-term storage conditions--80°C, Storage information--80°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Histone deacetylases commonly known as HDACs are enzymes that remove acetyl groups from histone proteins resulting in chromatin tightening and reduced gene transcription. These enzymes fall into several classes each with distinct structural features and typically have molecular masses ranging from 41 to 70 kDa. HDACs are expressed in various tissues including brain heart and muscle. HDAC activity is essential for the regulation of many cellular processes and can be measured using assays such as the HDAC activity assay or the colorimetric assay.
Biological function summary
HDACs play an important role in gene expression through the remodeling of chromatin structure. They are often part of multiprotein complexes with co-repressor proteins helping modulate the transcription of genes involved in cell cycle regulation and differentiation. This modulation impacts cellular activities such as proliferation and apoptosis making HDACs significant in normal cellular function and development. HDAC inhibitors serve as tools for research in studying HDAC functions and as potential therapeutic agents.
Pathways
HDACs influence the regulation of important cellular pathways like the cell cycle and apoptosis. They interact with other proteins such as histone deacetylase inhibitor proteins and transcription factors to regulate gene expression tightly. In the cell cycle pathway HDACs can repress the transcription of cyclin-dependent kinase inhibitors therefore promoting cell cycle progression. Their involvement in the apoptosis pathway is also important where they regulate pro-apoptotic and anti-apoptotic gene expression contributing to cellular homeostasis.
HDACs have a significant connection to cancer and neurological disorders. Overexpression or dysregulation of HDACs can lead to tumorigenesis by affecting cell cycle checkpoints and apoptotic pathways. HDAC inhibitors have been investigated as potential cancer therapies targeting cellular proliferation and survival mechanisms. In neurological disorders HDACs are associated with neurodegenerative diseases due to their role in neuronal gene expression regulation. Proteins such as the tumor suppressor p53 in cancer and tau proteins in Alzheimer's disease have connections to HDAC function linking these enzymes to pathological processes.
