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. It is also deeply involved in cellular proliferation, cell cycle and apoptosis.
Sample Type:
Human, mouse or rat. Tissue extract, cell lysate and nuclear extract.
Intended Use:
Applications include: Immunoprecipitation of HDAC2 complex from cell, nuclear, or tissue extract. Measurement of HDAC2 activity of immunoprecipitated complex/purified enzyme and screening for activators or inhibitors of HDAC2.
Test Principle:
The HDAC2 IP & Activity BioAssay(TM) Kit provides an antibody-based method to specifically immunoprecipitate the HDAC2 complex from cells & tissues and to measure HDAC2 activity fluorometrically. HDAC2 is immunoprecipitated from cell or nuclear extract(s) using HDAC2 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.
Kit Components:
HDAC Assay Buffer, 1x9ml
Cell Lysis Buffer, 1x100ml
HDAC Substrate, 1x100ul
Developer, 1x500ul
AFC (7-amino-4-trifluoromethyl coumarin) Standard, 1mM, 1x100ul
Rabbit HDAC2 Antibody, 1x250ul
Rabbit IgG (Control Antibody), 1x100ul
Protein A/G Sepharose Beads (50% slurry in 20% ethanol/H2O), 1x650ul
Positive Control (Jurkat Cell Lysate), lyophilized, 1x vial
Storage and Stability:
Store powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.