The EpiQuik™ Methyl-Histone H3K27 ChIP Kit is a convenient package of tools that allows the experimenter to investigate interactions of histone H3K27 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 H3K27 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 microwell 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 27 (H3K27) 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 H3K27 leads to chromatin gene silencing in many organisms. Methylated H3K27 has been involved in tumorigenesis through inactivation of tumor suppression genes. Thus, identification of genes silenced through H3K27 methylation is of particular significance for understanding and analyzing biological processes under the normal and pathological conditions such as cancer. 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 H3K27. ChIP coupled with microarrays could be further used for profiling or mapping histone H3K27 methylation patterns.
Principle & Procedure
This ChIP Kit contains all reagents required for carrying out a successful chromatin immunoprecipitation for methyl-histone H3K27 from mammalian cells. Particularly, this kit includes a ChIP-grade di-methyl histone H3K27 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 H3K27 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-Dimethyl-H3-K27 (1 mg/ml)*
8-Well Assay Strips (with Frame)
* For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.