The SensoLyte® p300 Assay Kit provides a convenient assay for the screening of enzyme inhibitors and for continuous measurement of p300 activity. For added convenience, this kit includes two peptide substrates: histone H3 (1-21) peptide and non-histone p53 peptide (368-386). After incubation with acetyl CoA and the substrate, the p300 enzyme generates acetylated H3 or p53 peptide and CoASH. The thiol groups of CoASH can be detected with a fluorogenic reagent at excitation/emission=389nm/513nm.
The SensoLyte® HAT (pCAF) Assay Kit provides a convenient assay for screening of enzyme inhibitors and for continuous measurement of pCAF activity. After incubation with acetyl CoA and histone H3 (1-21) peptide, the pCAF enzyme generates acetylated H3 peptide and CoASH. The detection of the CoASH thiol group is the same as above.
Histone acetyltransferases (HATs) enzymes regulate the acetylation of histones and non-histone proteins. The acetylation of the a-amino groups of lysine residues present at histone tails correlates largely with transcriptional activation, but it is also involved in DNA replication, DNA repair and protein–protein interactions. HATs play major roles in the control of cell fate and misregulation is implicated in the development of some human tumors. HAT p300 is a transcriptional coactivator that acetylates core histones facilitating chromatin decondensation and recruiting basic RNA polymerase machinery. Many non-histone proteins, such as p53, STATs, and alpha interferon receptor, also serve as substrates for p300. The p300/CBP-associated factor (pCAF) acetylates specific lysines on the N-terminal tails of histones H3 and H4.