The EpiQuik™ HDAC10 Assay Kit (Colorimetric) is a complete set of optimized buffers and reagents to colorimetrically measure the total amount of HDAC10 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.
Background InformationHistone 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. HDAC10 is a class II histone deacetylase containing 669 amino acid residues. HDAC10 contains a unique, putative second catalytic domain and resides in both the nucleus and cytoplasm. Data suggests that HDAC10 may play roles both in the nucleus, as a transcriptional modulator, and in the cytoplasm in an unidentified role. HDAC10 possesses histone deacetylase activity and represses transcription through interaction with many transcription factors. HDAC10 regulates many biological processes including cell cycle progression.
Principle & ProcedureIn 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 HDAC10 proteins contained in the sample bind to the substrate. The bound HDAC10 can be recognized with an HDAC10-specific antibody and 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 HDAC10 will be proportional to the intensity of the color development.
Fig. 1. Schematic procedure of the EpiQuik™ HDAC10 Assay Kit (Colorimetric).
Fig. 2. Illustrated standard curve generated with HDAC10 control.