The Flag-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified HDAC3 is greater than 90% homogeneous based on SDS-PAGE analysis.
Unit Definition (Activity)
1 unit equals 1 nanogram of purified protein.
Applications
Recombinant HDAC3 can be used 1) for in vitro function studies including transcription in the presence of co-factors suchas N-CoR; 2) for protein-protein interaction assay; and 3) for cell growth assay.
Formulation and Storage
The protein is in 20mM Tris-HCl pH7.9,100mM NaCl, 0.2mM EDTA, 1mM DTT and 20% glycerol. Stored at -70°C before use. Avoid repeated freeze thaw cycles.
The nucleosomal histones can be modified through reversible acetylation by histone acetyltransferases (HATs) and deacetylases (HDACs). Acetylation and deacetylation of histone proteins alters chromosome structure and affects transcription factor access to DNA. HDAC3 has an open reading frame of 428 amino acids. HDAC3 shares 53% amino acid identity with HDAC1 and 52% with HDAC2 (1). Observations indicate that class II HDACs regulate transcription by bridging the enzymatically active NCOR2-HDAC3 complex and select transcription factors. All HDACs were found to be ubiquitously expressed in immune and non-immune tissues. In human myeloid leukemia THP-1 cells, HDAC3 transfection resulted in increased size, aberrant nuclear morphology and cell cycle G2/M cell accumulation. Functional activity of the expressed HDAC3 protein was confirmed in alpha-HDAC3 antibody immunoprecipitates by a histone deacetylase assay. Study suggests the participation of HDACs in cell cycle progression and activation (2).
References
1. Yang, et al., (1997) J. Biol. Chem. 272: 28001-28007 2. Dangond,F., et al., (1998) Biochem. Biophys. Res. Commun. 242 (3): 648-652
DISCLAIMER
This products is recommended For RESEARCH USE ONLY and is Not qualified for Use in Diagnostic or Therapeutic Procedures.
