The EpiQuik™ Tri-Methyl-Histone H3K9 ChIP Kit is a convenient package of tools that allows the experimenter to investigate interactions of histone H3K9 tri-methylation and DNA in vivo efficiently. The entire procedure can be completed within 5 hours and produces far more superior results than any competitor kits. The EpiQuik™ Tri-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 (H3K9) is important for epigenetic regulation of the genes, as it can either be methylated or acetylated. It was well demonstrated that switching acetylation to methylation on H3K9 leads to chromatin gene silencing in many organisms. Methylated H3K9 has been involved in tumorigenesis through inactivation of tumor suppression genes. Tri-methylation of H3K9 is a stable heterochromatin mark, which promotes the gene silencing. Thus, identification of genes silenced through H3K9 tri-methylation is of particular significance for understanding and analyzing biological processes 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 H3K9. ChIP coupled with microarrays could be used further for profiling or mapping histone H3K9 methylation patterns.
Principle & Procedure
This ChIP kit includes all reagents required for carrying out a successful chromatin immunoprecipitation for tri-methyl-histone H3K9 from mammalian cells. Particularly, this kit includes a ChIP-grade tri-methyl-histone H3K9 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 tri-methyl-histone H3K9 protein-DNA complex, reversed, and purified through the specifically 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.
CP1 (Wash Buffer)
CP2 (Antibody Buffer)
CP3A (Pre-Lysis Buffer)
CP3B (Lysis Buffer)
CP4 (ChIP Dilution Buffer)
CP5 (DNA Release Buffer)
CP6 (Reverse Buffer)
CP7 (Binding Buffer)
CP8 (Elution Buffer)
Protease Inhibitor Cocktail (100X)*
Proteinase K (10 mg/ml)*
Normal Mouse IgG (0.5 mg/ml)*
Anti-Tri-Methyl-H3-K9 (1 mg/ml)*
8-Well Assay Strips (with Frame)
8-Well Strip Caps
* For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.