This article introduces the high-throughput analysis of epigenetic targets using the Agilent RapidFire/MS system, with a focus on the application of Sirtuin enzymes (SIRT). Sirtuin activity has been associated with inflammatory, cardiovascular, proliferative, neurodegenerative, and metabolic disorders. RapidFire/MS assays offer advantages over other assay formats, such as avoiding the use of radioactive substrates and the complications of secondary or coupled reactions. Additionally, direct measurement by mass spectrometry eliminates the need for fluorescent tags and enables detection of every modification state on a peptide.
The document details a method for studying sirtuins (SIRT1, SIRT2, and SIRT3) using Agilent RapidFire/MS systems. This approach offers advantages over traditional formats by circumventing the costs and special handling required for radioactive substrates and avoiding secondary reactions or fluorescent tags that can complicate data interpretation.
The system achieves a sustained throughput of seven seconds per sample.