The EpiQuik Total Histone Extraction Kit is a complete set of buffers and reagents optimized for extracting total core histone proteins (H2A, H2B, H3, and H4) from mammalian cells or tissues in a simple 60-minute procedure. Post-translational modifications (PTMs) in histone extracts remain intact and therefore can be used with Epigentek’s histone modification assay kits or in a variety of downstream applications for methylation, acetylation, phosphorylation, sumoylation, ubiquitination, citrullination, and ADP of histones. ribosylation studies.
- Simple and pre-optimized 1-hour protocol.
- Conveniently includes all essential reagents to perform histone extraction.
- A standardized procedure for reproducible results.
- Extracts a high yield of total core histones from as little as 1 mg of tissues.
- Post-translation modifications remain intact.
- It does not affect the state or levels of histone modification.
Histones are the main protein components of chromatin in biology. They act like spools around which DNA is wound and also play a role in gene regulation. The core histones include H2A, H2B, H3, and H4. Histones undergo post-translational modifications, which alter their interaction with DNA and nuclear proteins. Histones H3 and H4 have long tails that protrude from the nucleosome, which can be covalently modified in various places (ie, methylation, acetylation, phosphorylation, etc.). Histone modifications act in various biological processes such as gene regulation, DNA repair, and chromosomal condensation (mitosis). Combinations of modifications are believed to constitute a “histone code” and understanding this code can help answer questions about human development and disease.
Principle and procedure
The EpiQuik Total Histone Extraction Kit simply applies our proprietary histone isolation buffers to cell or tissue samples. After treatment with pre-lysis, lysis, and equilibrium buffers, the total core histones are easily extracted for immediate use or storage under suitable conditions.
Materials to get started
The minimum amount of starting materials can be as low as 10 5 cells or 1 mg of tissue. For best results, the number of cells should be greater than 106 cells or the amount of tissue should be greater than 10 mg. A total of 100 standard extractions can be performed with this kit (using 107 cells or 100 mg of tissues per extraction). The yield of total histone proteins is approximately 0.4 mg per 107 cells or 100 mg of tissue. Performance may vary based on cell or tissue type.