In the last decade, mass spectrometry (MS) has become the method of choice to profile histone PTMs in a systematic manner. Indeed, MS analysis does not require any a priori knowledge of the modification site, offers a comprehensive view of multiple histone PTMs and their combinations, and provides accurate quantification of even mild relative changes among samples. To meet the downstream applications such as MS-based histone PTM assays, Creative Proteomics has adapted and developed a series of methods to extract and enrich histones from different types of samples while preserving histone PTMs, including cell culture, tissue, and primary cells. We are dedicated to providing our global customers with the fastest, most efficient, most affordable service to support their projects.
Overview of histones
Histones are small hydrophilic proteins responsible for DNA compaction in the nucleus and transcriptional regulation of basic genes, which is realized by a large number of combinatorial modifications, mainly in its N-terminal tail. Histones consist of histone H3, H4, H2A, and H2B, which combine to form the basic unit of chromatin, nucleosome, and linker histone H1. PTMs have been found on more than 60 amino acid residues at different sites of histone sequence, and acetylation and methylation are the most studied and characterized in these different types of modifications. Alterations in the levels of histone PTMs as well as aberrant expression or mutations in the enzymes are related to their deposition and removal, leading to defects in cellular processes and may result in different diseases, especially cancer.
The service offering at Creative Proteomics
- Histone extraction
We are dedicated to providing fast, efficient, and affordable histone extraction and purification service for cell culture, tissue, and primary cells while preserving histone PTMs. Since histones are very basic proteins, a typical histone separation scheme takes advantage of their hydrophilicity and solubility in strong acids. During the process of histone extraction, we add inhibitors to help preserve histone modifications (such as acetylation, methylation, and phosphorylation) and use optimized protocols for efficient recovery of core histones (including histone H3, H4, H2A , and H2B). - Histone purification
Starting with well-established protocols to extract histones from cells, each protocol has been optimized to address the limitations associated with specific samples. Then, our histone purification service uses reversed-phase high-performance liquid chromatography (RP-HPLC), hydrophilic-interaction chromatography (HILIC), and precipitation steps for histone purification. - Histone modified peptide enrichment
To accelerate the success of our customers' projects related to histone PTM analysis, we offer a range of commonly used separation and modified peptide enrichment strategies for customers to choose from, including immobilized metal affinity chromatography (IMAC), titanium dioxide (TiO2)-based phosphopeptide enrichment, and so on.
Applications of our histone enrichment and purification service
Our service can achieve the isolation/enrichment of histones from different types of primary samples, including
- Histone isolation/enrichment from frozen tissues.
- Histone isolation/enrichment from cell culture.
- Histone isolation/enrichment from primary cells.
Creative Proteomics is dedicated to the comprehensive and reliable identification and quantification of histone PTMs. Our PTM proteomics analysis platform has a team of experts and state-of-the-art equipment to help you navigate successful research projects through strong workflow, powerful analysis, and tailored services. If you are interested in our services, or just would like to share some feedback, please contact us. We would love to hear from you.
References
- Noberini, Roberta, et al. "Enrichment of histones from patient samples for mass spectrometry-based analysis of post-translational modifications." Methods 184 (2020): 19-28.
- Lu, Congcong, et al. "Accelerating the Field of Epigenetic Histone Modification Through Mass Spectrometry–Based Approaches." Molecular & Cellular Proteomics 20 (2021).
Our products and services are for research use only.
