The EpiQuik™ Chromatin Immunoprecipitation (ChIP) Kit is a convenient package of tools that allows the experimenter to perform chromatin immunoprecipitation (ChIP) 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. 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™ 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 microwell 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
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)
CP3A (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)*
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 firstname.lastname@example.org along with your contact information and institution name.
References & Citations
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Cheetham, S. et al. (Jun 2008). SPARC promoter hypermethylation in colorectal cancers can be reversed by 5-Aza-2'deoxycytidine to increase SPARC expression and improve therapy response. British Journal of Cancer 98(11): 1810-9. PubMed Abstract
Yamada N. et al. (Apr 2008). MUC1 expression is regulated by DNA methylation and histone H3 lysine 9 modification in cancer cells. Cancer Res. 68(8): 2708-16. PubMed Abstract
Ulleras, Erik et al. (Mar 2008). NFAT but not NF-κB is critical for transcriptional induction of the pro-survival gene A1 following IgE receptor activation in mast cells. Blood 111(6): 3081-9 PubMed Abstract
Venteclef, Nicolas et al. (Feb 2008). Regulation of anti-atherogenic apolipoprotein M gene expression by the orphan nuclear receptor LRH-1. J. Biol. Chem. 283(7): 3694-701. PubMed Abstract
Liu, DX et al. (Nov 2007). Transcriptional activation of p53 by Pitx1. Cell Death & Differentiation 14(11): 1893-907. PubMed Abstract
Teng, Yun et al. (Nov 2007). AS1411 alters the localization of a complex containing protein arginine methyltransferase 5 and nucleolin. Cancer Research 67(21): 10491-500. PubMed Abstract
Sakurai, Takuya et al. (Nov 2007). Functional roles of Fli-1, a member of the Ets family of transcription factors, in human breast malignancy. Cancer Science 98(11): 1775-84. PubMed Abstract
Mora-Lopez, Francisco et al. (Nov 2007). Human BSAP and BLIMP1 conform an autoregulatory feedback loop. Blood 110(9): 3150-7 PubMed Abstract
Gao, Song et al. (Jun 27, 2007). Cloning and characterization of the rat lysyl oxidase genepromoter: Identification of core promoter elements andfunctional nuclear factor I binding sites.J. Biol. Chem. 282(35): 25322-37. PubMed Abstract
Venteclef, Nicolas et al. (Feb 2007). Interleukin-1 receptor antagonist induction as an additional mechanism for liver receptor homolog-1 to negatively regulate the hepatic acute phase response. J. Biol. Chem. 282(7): 4393-9. PubMed Abstract