The ChromaFlash™ One-Step Magnetic ChIP Kit is a complete set of optimized buffers and reagents, using a magnetic beads-based method, to carry out a successful chromatin immunoprecipitation for the selective enrichment of a chromatin fraction containing specific DNA sequences from isolated chromatin, in order to investigate the interaction between proteins and DNA in a cell.
This kit can be used with the EpiSonic™ Multi-Functional Bioprocessor 1000 sonication device, or as a standalone kit*. With the EpiSonic, chromatin shearing and immunoprecipitation steps are optimally processed at the same time, reducing the entire procedure (from intact chromatin sample to ready- for-use DNA) to less than 70 minutes.
* Standalone protocol without the EpiSonic 1000 is a 4 hour procedure.
Next generation sequencing and microarrays are becoming major technologies of massive protein-DNA analysis. The major features of these technologies are their rapidness and high throughput capabilities. To be compatible with these new technologies, rapid and massive generation of target protein-bound DNA is critically required. To meet this requirement, Epigentek developed ChromaFlash™, a new ChIP technology, and incorporated it into the ChromaFlash™ Magnetic One-Step ChIP Kit. By utilizing this kit and the powerful EpiSonic™ Multi-Functional Bioprocessor 1000, chromatin shearing and immunoprecipitation can be simultaneously processed, which greatly propels ChIP to the fastest speeds in a high throughput format with much greater efficiency. The kit has the following advantages and features:
- The fastest and most convenient ChIP method. The entire procedure (from intact chromatin sample to ready-for-use DNA) is less than 70 minutes with the actual handling time being less than 15 minutes due to simultaneous processing of chromatin shearing and immunoprecipitation ("One-Step ChIP").
- 96-well plate format makes the assay flexible. Either (a) manual with one single reaction each time; or (b) high throughput with 96 reactions each time.
- Highly efficient enrichment. Enrichment ratio of positive to negative control > 200, and an extremely low number of cells required (as low as 5,000 cells per ChIP reaction).
- High reproducibility. Pre-optimized ChIP conditions and, with the EpiSonic 1000, digitally acoustic- controlled reaction processing in sealed vessels make the ChIP procedure consistent.
- Wide downstream analysis compatibility. Compatible with various downstream analysis workflows including ChIP-PCR, ChIP-on-chip, and ChIP-seq.
See also a quick chart to compare ChIP kits.
Principle & Procedure
The ChromaFlash™ One-Step Magnetic ChIP Kit contains all necessary reagents required for carrying out a successful chromatin immunoprecipitation directly from chromatin extracts isolated from mammalian cells or tissues. This kit includes a positive control antibody (RNA polymerase II), a negative control non-immune 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 non-immune IgG. Immunoprecipitated DNA is then cleaned, released, and eluted. Eluted DNA can be used for various downstream applications such as ChIP-PCR, ChIP-on-chip, and ChIP-seq.
Accelerated ChIP Protocol with ChromaFlash...
Using this kit's accelerated ChIP protocol, chromatin shearing and immunoprecipitation are simultaneously processed with EpiSonic's digitally programmed acoustics and pre-optimized reaction conditions, enabling the ChIP to be completed in less than 60 minutes with a high efficiency through an accelerated and maximized antibody-protein/DNA interaction.
...or Standalone ChIP Protocol with ChromaFlash
Alternatively, this kit also includes a standalone protocol that does not require any special equipment. You can use your own sheared chromatin with this protocol for a ChIP procedure that can be completely finished in less than 4 hours.
Fig. 1. Schematic procedure for using the accelerated protocol of the ChromaFlash™ One-Step Magnetic ChIP Kit.
Fig. 2. Analysis of enrichment of RNA polymerase II in GAPDH and MLH1 promoters by the ChromaFlash™ One-Step Magnetic ChIP Kit with chromatin extract prepared from formaldehyde fixed colon cancer cells. Captured DNA was used for analyzing levels of RPII enriched in the GAPDH and MLH1 promoters.