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David E. Volk,
Ganesh L.-R. Lokesh,
Alpa M. Nick,
Anil K. Sood,
Kevin P. Rosenblatt*,
and David G. Gorenstein,
Several assay methods have been developed for quantifying the activity of histone deacetylases (HDACs and sirtuins), histone methyltransferases (HMTs), and histone demethylases (HDMs). These include radioactive assays, enzymelinked immunoassays (ELISA), mass spectrometry, and enzyme-coupled detection of fluorescent peptides or reaction co-products (e.g.
Post-translational modifications of histone proteins play an important part in a wide array of cellular processes including regulation of gene transcription, DNA repair, cell cycle, and metabolism control. For instance, transcriptional activation is associated with acetylation of Histone H3 on residues K9 and K14, and methylation on K4.
In eukaryotes, the covalent modification of histones has a crucial role in chromatin architecture and plays an important part in a plethora of cellular processes, from chromatin remodeling and transcriptional regulation, to DNA repair and cell cycle control. While histone acetylation is generally associated to an open chromatin state and transcriptional activation, methylation of histones has been related to either activating or repressive functions.
The PressureBioSciences Inc.(PBI) focuses on development of specialized products for biological sample preparation which utilize alternating ultra-high hydrostatic pressure–pressure cycling technology(PCT). PCT is a fundament althermo dynamic process which destabilizes molecular interactions by rapidly and repeatedly raising and lowering pressure in the action vessel from atmospheric to levels of up to 45,000 psi.
The Strep-tag II is ideal as an affinity tag due to the small size (8 aa, 1 kDa), often making removal of the tag unnecessary.
StrepTrap™ HP 1 ml and 5 ml columns have been specially developed for high specificity towards the Strep-tag II, with the prepacked format facilitating fast and convenient purification.
Protein kinases regulate many different signaling pathways and cellular processes. They have been implicated in various cancers, typically displaying increased activation, and are now being investigated as potential therapeutic targets.
Maltose binding protein (MBP) is beneficial as an affinity tag due to its ability to increase expression level and solubility of the fusion protein. The MBPTrap™ HP 1 ml and 5 ml columns have been specially developed for high specificity towards the MBP-tag and the prepacked format facilitate fast, convenient and reproducible purifications.
Gel filtration (GF) is an excellent tool to acquire information about sizing (identity), purity and specially the multimeric state of a protein of interest. Superdex™ 200 and Superdex 75 are supreme GF media for such analysis.