The EpiQuik™ Methyl-Histone H3K9 ChIP Kit is a convenient package of tools that allows the experimenter to investigate interactions of histone H3K9 methylation and DNA in vivo efficiently. The entire procedure can be completed within 5 hours and produces far superior results than any competitor kits. The EpiQuik™ Methyl-Histone H3K9 ChIP Kit is suitable for combining the specificity of immunoprecipitation with qualitative and quantitative PCR, ChIP-Seq, and ChIP-on-chip. This kit has the following advantages:
- The fastest procedure available, which can be finished within 5 hours.
- Strip microplate format makes the assay flexible: manual or high throughput.
- Columns for DNA purification are included: save time and reduce labor.
- Compatible with all DNA amplification-based approaches.
- Simple, reliable, and consistent assay conditions.
The position of the histone H3 at lysine 9 (H3-K9) is important for epigenetic regulation of the genes, as it can be either methylated or acetylated. It was well demonstrated that switching acetylation to methylation on H3-K9 leads to chromatin gene silencing in many organisms. Methylated H3-K9 has been involved in tumorigenesis through inactivation of tumor suppression genes. Thus, identification of genes silenced through H3-K9 methylation is of particular significance for understanding and analyzing biological process under the normal and pathological conditions. Chromatin Immunoprecipitation (ChIP) is a powerful technique for studying protein-DNA interaction in vivo. ChIP also offers an advantageous tool that allows identification of silenced genes associated with methylated histone H3-K9. ChIP coupled with microarrays could be further used for profiling or mapping histone H3-K9 methylation patterns.
Principle & Procedure
This ChIP kit includes all reagents required for carrying out a successful chromatin immunoprecipitation for methyl-histone H3-K9 from mammalian cells. Particularly, this kit includes a ChIP-grade di-methyl histone H3-K9 antibody and a negative control normal mouse IgG. Chromatin from the cells is extracted, sheared, and added into the microwell immobilized with the antibody. DNA is released from the antibody-captured methyl-histone H3-K9 protein-DNA complex, reversed and purified through the specially designed F-Spin Column. Eluted DNA can be used for various down-stream applications.
Starting materials can include various cell samples. In general, the input amount should be from 1,000,000 to 4,000,000 cells for each reaction.