Acetylated histones are recombinant or chemically/enzymatically modified histone proteins bearing acetylation at one or more defined lysine residues (e.g., H3K9ac, H3K14ac, H3K27ac, H4K5ac, H4K8ac, H4K16ac). They serve as precise reagents for epigenetics research, chromatin biochemistry, antibody validation, enzymatic assays, chromatin assembly, and structural studies. Defined acetylation states enable mechanistic dissection of histone acetylation effects on nucleosome structure, reader binding, transcriptional activation, and enzymatic regulation.

Key Features

• Site-Specific Acetylation: Single-site or multisite acetylated histones with defined modification stoichiometry (e.g., H3K27ac mono-acetylated; H3K9acK14ac double-acetylated).
• Multiple Production Methods: Chemical ligation (native chemical ligation or expressed protein ligation), enzymatic acetylation by histone acetyltransferases (HATs), or amber codon incorporation of acetyl-lysine analogs, selected according to the modification site, complexity, and requirement for native linkage.
• Available Formats: Single recombinant histone proteins, pre-assembled histone octamers containing acetylated subunits, mononucleosomes, nucleosome arrays with defined acetylation patterns, and peptide controls.
• High Purity and Characterization: Purity typically exceeds 90–95%, verified by SDS-PAGE and HPLC, while site occupancy and molecular mass are confirmed by LC-MS/MS.
• Customization Options: Choice of species (human, mouse, chicken), histone variant backbone (H3.1, H3.3), combined post-translational modifications (acetylation with methylation and/or phosphorylation), affinity tags (His, FLAG, Avi), and fluorescent or biochemical labels for downstream applications.

Common Applications

• Antibody Validation: Gold-standard controls for assessing the specificity of acetyl-lysine-specific antibodies in Western blotting, dot blotting, peptide array analysis, and chromatin immunoprecipitation (ChIP) validation.
• Reader Domain Binding Assays: Investigation of bromodomain and other acetyl-lysine reader protein interactions using biophysical techniques such as isothermal titration calorimetry (ITC), surface plasmon resonance (SPR), fluorescence polarization (FP), and inhibitor screening assays.
• Enzyme Activity Assays: Use as substrates or controls in histone deacetylase (HDAC) and sirtuin activity assays, as well as in histone acetyltransferase (HAT) activity and kinetic studies.
• Nucleosome Reconstitution: Evaluation of how site-specific acetylation influences nucleosome stability, chromatin accessibility, nucleosome remodeling, and higher-order chromatin organization.
• Structural Biology Studies: Generation of homogeneous and precisely modified nucleosomes for cryo-electron microscopy (cryo-EM), X-ray crystallography, and structural analysis of chromatin-associated complexes.
• Assay Development and High-Throughput Screening: Application as substrates or positive and negative controls in screening campaigns aimed at identifying epigenetic modulators and chromatin-targeting compounds.
• Mass Spectrometry Standards: Utilization of acetylated histones and peptides as quantitative standards and reference materials for proteomics method development and validation.