The EpiQuik™ Tissue Chromatin Immunoprecipitation (ChIP) Kit is a convenient package of tools that allows the experimenter to perform chromatin immunoprecipitation (ChIP) from mammalian tissues at extraordinarily rapid speeds and consistency, superior to all other current ChIP methods available. The kit is ready-to-use and provides all the essential components needed to carry out a successful ChIP experiment on tissues. The EpiQuik™ ChIP kits are suitable for combining the specificity of immunoprecipitation with qualitative and quantitative PCR, MS-PCR, DNA sequencing, and southern blot as well as DNA microarray.
WHY CHOOSE THE EPIQUIK™ TISSUE CHROMATIN IMMUNOPRECIPITATION CHIP KIT?
- Straightforwardly put: It is the truly number one fastest procedure available. The entire procedure can be completed within an amazingly short 5 hour period with superb results.
- The ChIP procedure has been drastically simplified -- extremely short and easy to follow.
- Strip microplate format makes the assay flexible: manual or high throughput.
- Columns for DNA purification are included: save tremendous amounts of time and reduce unnecessary physical labor.
- Compatible with all DNA amplification-based approaches.
- Achieves very reliable and consistent assay conditions.
Principle & Procedure
The EpiQuik™ Tissue Chromatin Immunoprecipitation kit contains all reagents required for carrying out a successful chromatin immunoprecipitation from mammalian tissues. Particularly, this kit includes a positive control antibody (RNA polymerase II), a negative control normal mouse IgG, and GAPDH primers that can be used as a positive control to demonstrate the efficacy of the kit reagents and protocol. RNA polymerase II is considered to be enriched in the GAPDH gene promoter that is expected to be undergoing transcription in most growing mammalian cells and can be immunoprecipitated by RNA polymerase II but not by normal mouse IgG.
In this ChIP, cells are cross-linked with formaldehyde and chromatin is extracted. The chromatin is then sheared and added into the microwell immobilized with affinity antibodies. Cross-linked DNA is released from antibody-captured protein-DNA complex, reversed and purified through the specifically designed Fast-Spin Column. Eluted DNA can be used for various down-stream applications.
CP1 (wash buffer)
CP2 (antibody buffer)
CP3 (lysis buffer)
CP4 (CHIP dilution buffer)
CP5 (DNA release buffer)
CP6 (reverse buffer)
CP7 (binding buffer)
CP8 (elution buffer)
Protease inhibitor cocktail (100x)*
Normal mouse IgG (1 mg/ml)*
Anti-RNA polymerase II (1mg/ml)*
Proteinase K (10 mg/ml)*
Control primer (GAPDH)
Forward (20 µM)*
Reverse (20 µM)*
8-well assay strips (with frame)
8-well strip caps
* Spin the solution down to the bottom before use.
User Guide & MSDS
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing email@example.com along with your contact information and institution name.
References & Citations
Fuchikami, Manabu E. et al. (Jun 2008). Single immobilization stress differentially alters the expression profile of transcripts of the brain-derived neurotrophic factor (BDNF) gene and histone acetylation at its promoters in the rat hippocampus. Int J Neuropsychopharmacol Epub ahead of print PubMed Abstract
O'Connor JC. et al. (May 2008). A novel antioxidant function for the tumour suppressor gene p53 in the retinal ganglion cell. Invest Ophthalmol Vis Sci Epub ahead of print PubMed Abstract
Aoyama, Tomoki et al. (Jan 2008). Cell-specific epigenetic regulation of ChM-I gene expression: Crosstalk between DNA methylation and histone acetylation. Biochem Biophys Res Commun. 365(1): 124-30. PubMed Abstract
Johnson, Carrie E. et al. (Dec 2007). Differential Apaf-1 levels allow cytochrome c to induce apoptosis in brain tumors but not in normal neural tissues. Proceedings of the National Academy of Sciences of the USA 104(52): 20820-5. PubMed Abstract