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Histone Modifications

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SIRTainty™ Assay Principle


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Sirtuin-mediated deacetylation of unlabeled peptide substrate generates nicotinamide as a product. The SIRTainty™ assay couples sirtuin enzyme activity to nicotinamidase, which cleaves nicotinamide into nicotinic acid and free ammonia. A developer reagent is added, which reacts with the free ammonia to generate a fluorophore. The resulting fluorescent signal is quantified with a conventional fluorometric plate reader.

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Regulation of Chromatin by Histone Modifications

A nucleosome consists of an octamer of histone proteins tightly associated with DNA. Accessibility to regional DNA sequences is regulated by post-translational modifications to these histone proteins.Histone modifications regulate DNA transcription, repair, recombination, and replication, and can alter local chromatin architecture. The most commonly studied and best understood histone modifications are acetylation, phosphorylation, methylation, and ubiquitination.

Merck offers over 200 validated antibodies, recombinant proteins, and kits to analyze histone modifications.

Histone Deacetylase (HDAC)

SIRTainty™ Class III HDAC Assay
Sirtuins (class III HDACs) became the focus of intense research when it was discovered that their activation led to reduced incidence of aging and age-related diseases, including diabetes.

The SIRTainty™ Assay Features
  • Sensitive: Almost five times more sensitive than assays dependent on labeled substrates.
  • Flexible: Analyze multiple sirtuin isoforms using virtually any substrate.
  • Reliable: Avoid fluorophore-mediated activation by compounds such as resveratrol.
  • Fast and Easy: Homogeneous, no-wash, 96-well assay minimizes hands-on time.
DescriptionCatalogue No.
SIRTainty™ Class III HDAC Assay 17-10090
SIRT1 Deacetylase 17-370
HDAC Assay Kit, Colorimetric 17-374
HDAC Assay Kit, Fluorometric 17-356

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Sirt1, Sirt2, and Sirt3 exhibit preference for acetylated vs. nonacetylated peptides.