The EpiQuik™ Total Histone Extraction Kit is a complete set of optimized buffers and reagents for extracting total core histone proteins (H2A, H2B, H3, and H4) from mammalian cells or tissues in a simple 60 minute procedure. The post-translational modifications in the histone extracts are kept intact and thus can be used with Epigentek's histone modification assay kits or in a variety of downstream applications for histone methylation, acetylation, phosphorylation, sumoylation, ubiquitination, citrullination, and ADP-ribosylation studies.
- Pre-optimized and simple 1 hour protocol.
- Conveniently includes all essential reagents to carry out a histone extraction.
- Standardized procedure for reproducible results.
- Extracts a high yield of total core histones from as little as 1 mg of tissues.
- Post-translational modifications are kept intact.
- Does not affect histone modification status or levels.
Histones are the chief protein components of chromatin in biology. They act as spools around which DNA winds, 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. The H3 and H4 histones have long tails protruding from the nucleosome, which can be covalently modified at several places (i.e., methylation, acetylation, phosphorylation, etc.). Histone modifications act in diverse biological processes such as gene regulation, DNA repair and chromosome condensation (mitosis). Combinations of modifications are thought to constitute a "histone code" and an understanding of this code may help to answer questions about human development and disease.
Principle & 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 balance buffers, the total core histones are easily extracted for immediate use or storage under proper conditions.
The minimal amount of starting materials can be as low as 105 cells or 1 mg tissue. For the best results, the cell number should be greater than 106 cells or the tissue amount should be greater than 10 mg. A total of 100 standard extractions (using 107 cells or 100 mg tissues per extraction) can be performed with this kit. Yield of the total histone proteins is approximately 0.4 mg per 107 cells or 100 mg of tissue. The yield may vary depending on cell or tissue type.
Fig. 1. Schematic procedure of chromation isolation with the EpiQuik™ Total Histone Extraction Kit.
Fig. 2. Histone extracts were prepared from MCF-7 cells using the EpiQuik Total Histone Extraction Kit and acetyl histone H3-K9 was quantified using the EpiQuik Global Acetylated Histone H3-K9 Quantification Kit (Fluorometric) (Cat. No. P-4011).