The EpiQuik™ HDAC3 Assay Kit (Colorimetric) is a complete set of optimized buffers and reagents to colorimetrically measure the total amount of HDAC3 proteins quantitatively, through an ELISA-like method, from fresh tissue and cultured cells of human and mouse without the need for electrophoresis and transfer processes.
- Very fast procedure, which can be finished within 4 hours.
- Innovative colorimetric assay to quantitatively measure HDAC3 protein amount without the need for electrophoresis.
- High sensitivity and specificity -- HDAC3-specific detection with a detection limit as low as 0.2 ng of HDAC3 protein.
- Strip microplate format makes the assay flexible: manual or high throughput analysis.
- HDAC3 control is included, which allows for the HDAC3 protein amount of the sample to be properly quantified.
- Simple, reliable, and consistent assay conditions.
Histone deacetylases (HDACs) play a critical role in transcriptional repression of gene expression in eukaryotic cells by catalyzing the hydrolytic removal of acetyl groups from histone lysine residues. HDACs are tightly involved in cell cycle regulation, cell proliferation, and the development of human cancer. HDAC inhibition displays significant effects on apoptosis, cell cycle arrest, and differentiation in cancer cells. HDAC inhibitors are currently being developed as potential anti-cancer agents. Three distinct families of HDACs have been described, comprising a group of at least 20 proteins in humans. HDAC3 is a class I histone deacetylase containing 428 amino acid residues. HDAC3 acts on chromatin via the formation of large multiprotein complexes that affect gene transcription and correlate with epigenetic repression. Also, through interaction with numerous transcription factors, HDAC3 regulates many biological processes including cell cycle progression. HDAC3 is the most highly expressed class I HDAC throughout the brain, including the hippocampus, and is a critical negative regulator of long-term memory formation. HDAC3 is also overexpressed in many different cancer types such as colon cancer and breast cancer.
Principle & Procedure
In an assay with this kit, the unique HDAC affinity substrate is stably coated on the strip well. The sample is added into the well and HDAC3 proteins contained in the sample bind to the substrate. The bound HDAC3 can be recognized with an HDAC3-specific antibody and can be colorimetrically quantified through an ELISA-like reaction. After reading the optical density on a microplate spectrophotometer at a wavelength of 450 nm, the amount of HDAC3 will be proportional to the intensity of the color development.
Fig. 1. Schematic procedure of the EpiQuik™ HDAC3 Assay Kit (Colorimetric).
Fig. 2. Illustrated standard curve generated with HDAC3 control.
Fig. 3.Nuclear extracts were prepared from HeLa cells and the ODs generated from HDAC3 are measured.