CBP and p300 are large nuclear proteins that bind to many sequence-specific factors involved in cell growth and/or differentiation, including c-jun and the adenoviral oncoprotein E1A. The protein encoded by this gene associates with p300/CBP. It has in vitro and in vivo binding activity with CBP and p300, and competes with E1A for binding sites in p300/CBP. PCAF belongs to the GCN5/PCAF family of nuclear HATs. The proteins consist of N-terminal and C-terminal domains separated by a deep hydrophobic cleft. The C-terminus contains the HAT domain and a bromodomain, while the N-terminus contains a PCAF-specific domain (2). Numerous studies indicate that these HATs function as histone- acetylating transcriptional co-activators (3-5). Besides histones, PCAF also acetylates non-histone proteins (2, 6).It has histone acetyl transferase activity with core histones and nucleosome core particles, indicating that this protein plays a direct role in transcriptional regulation.
Source: The PCAF was expressed in baculovirus system and purified by an affinity column in combination with FPLC chromatography.
Applications: Recombinant PCAF can be used 1) for protein-protein interaction assay; 2) for in vitro transcription assay; 3) for in vitro acetylation assay; and 4) for cell growth assay.
Quality Control: Purified protein is greater than 95% homogeneous and contains no detectable proteases, DNase and RNase activity.
Unit Definition: 1 unit equals 1 nanogram of purified protein. 1-5 units are sufficient for a gel mobility shift assay in a 20µl reaction; 50-100 units are sufficient for reconstituted transcription assay and 100-200 units are sufficient for a protein-protein interaction assay or an acetylation assay.
Concentration: 0.5 mg/ml (in 1x dilution buffer A with 500mM KCl)
Reagents Supplied: 1x dilution buffer A: 20 mM Tris-Cl (pH 8.0), 20% Glycerol, 100 mM KCl, 1 mM DTT and 0.2 mM EDTA.
