Histone acetylation, including H3 and H4, is performed through the addition of an acetyl group to lysine residues in the N-terminal tail and to the surface of the nucleosome core of histone proteins, which has long been associated with the regulation of chromatin structure and the recruitment of transcription factors to gene promoters, and is tightly involved in cell cycle regulation, cell proliferation and apoptosis. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) are critical in controlling H3 acetylation. Acetylated and deacetylated histones within chromatin are recognized as epigenetic tags by loosening (euchromatin) or compacting (heterochromatin) chromatin structure, subsequently increasing or decreasing gene transcription levels. Quantification of acetylated histone would provide meaningful information for further characterizing the acetylation patterns or sites.

Creative BioMart is a leading professional organization in offering acetylated histone quantification service with high efficiency, consistency, and convenience, and is specialized in all the essential procedures of detection to carry out a successful assay, including rapid tissue digestion, cell lysis, and histone extraction, based on ELISA methods. Highly effective antibodies are applied to enrich acetylated peptides with the purpose of achieving large-scale acetylation quantification.

What Is Acetylated Histone Quantification (AHQ) Assay?

Histone acetylation at specific lysines has been involved in pathological processes such as cancer or lupus progress. AHQ assay is designed for measuring the amount of total acetylated H3 or H4 or acetylation levels of modified histones at particular lysines in plasma or serum by employing an ELISA-based colorimetry/fluorometry. Quantification of acetylated levels of overall histone or specific lysines would give a better elucidation about epigenetic regulation of gene activation and silencing, histone modification-associated pathological processes, screening of disease-related biomarkers, and exploration of drugs targeting HATs or HDACs. Biological fluid samples such as plasma and serum and mammalian cells from human, mouse or rat could be applied in AHQ assay.

Procedure of AHQ Assay

In AHQ assay, the acetylated histones in prepared sample are captured on the strip wells that are coated with corresponding antibody, and then can be detected with another labeled antibody. Ultimately, a color development reagent is added to produce color or fluorescence, whose absorption intensity is proportional to the ratio of acetylated histones. The absolute amount of histone can be quantitated by comparing to the standard control.

Figure 1. Workflow of AHQ assay.

Advantages of Our AHQ Service

1. Simple, reliable, and repeatable testing conditions
2. Quick and efficient procedure: Finished within a few hours
3. Creative colorimetric or fluorometric methods: No need for radioactivity, electrophoresis or chromatography
4. Excellent detection sensitivity and specificity: Controls are conveniently included
5. Strip microwell format: Flexible for manual or high throughput analysis

Creative BioMart has utilized the power of Elisa-based chromometry to develop a high-throughput approach for the quantification technology of acetylated histone at low-cost in short time. Supported by extensive experience and advanced platform, we are confident to provide fully customizable services in consultation with clients with the aim to satisfy each demand from our customers. Please feel welcome to contact us for any problem about your program in AHQ and we are pleased to give you complementary technical consultation.

Reference
Verdone, L.; et al. Histone acetylation in gene regulation. Brief Funct Genomi. 2006, 5(3): 209–221.