The ChromaFlash™ Plant Chromatin Extraction Kit is a complete set of optimized buffers and reagents for isolating chromatin or DNA-protein complexes from plants in a simple and rapid format. Chromatin prepared by this kit can be used in a variety of chromatin immunoprecipitation methods. It is also the recommended method for obtaining chromatin required by Epigentek’s one-hour ChIP method using the ChromaFlash™ One-Step ChIP Kit or the ChromaFlash™ One-Step Magnetic ChIP Kit. The isolated chromatin can also be used in other chromatin-related applications such as in vitro protein-DNA binding assays, or nuclear enzyme assays.
- Extremely fast procedure: the entire procedure from plant samples to ready-to-use chromatin is less than 2 hours.
- Convenient and flexible: the kit is suitable for preparing both native chromatin and cross-linked chromatin from monolayer or suspension cells, or from tissues.
- Unsheared chromatin makes it customizable for various analysis workflows that require either intact or fragmented chromatin, including ChIP, in vitro protein-DNA interaction analysis, nuclear enzyme assays, etc.
Chromatin immunoprecipitation (ChIP) offers an advantageous tool for studying protein-DNA interaction. With ChIP, the experimenter can determine if a specific protein binds to the specific sequences of a gene in living cells by combining it with PCR (ChIP-PCR), microarray (ChIP-chip), or sequencing (ChIP-Seq) techniques. For example, the measurement of the amount of methylated histone H3 at lysine 9 (meH3-K9) associated with a specific gene promoter region under various conditions can be achieved through a ChIP-PCR assay, while recruitment of meH3-K9 to the promoters on a genome-wide scale can be detected by ChIP-chip. In particular, a ChIP method with specific antibodies directly against various transcriptional factors is widely in demand.
Principle & Procedure
The ChromaFlash™ Plant Chromatin Extraction Kit contains all the reagents required for carrying out a successful chromatin extraction directly from plants. Cell membranes of the sample, with or without cross-linking, are broken down using the provided lysis buffer. Chromatin or DNA-protein complex is then extracted with the extraction buffer. The extracted chromatin can then be diluted with chromatin buffer and stored at the appropriate temperature.
Fig. 1. Schematic procedure of the ChromaFlash™ Plant Chromatin Extraction Kit.
Fig. 2. ChIP analysis of tri-methyl H3-K27 enriched in AGAMOUS gene with chromatin extract prepared from 2-week-old icu2-1/icu2-1 seedlings using the ChromaFlash™ Plant Chromatin Extraction Kit.