ChromaFlash Chromatin Extraction Kit
The ChromaFlash ™ Chromatin Extraction Kit is a complete set of buffers and reagents optimized for isolating chromatin or DNA-protein complex from mammalian cells or tissues in a simple and rapid format. Chromatin prepared with this kit can be used in a variety of chromatin immunoprecipitation methods. It is also the recommended method of obtaining the chromatin required by the Epigentek One-Hour ChIP method using the ChIP ChromaFlash ™ One-Step Kit. Isolated chromatin can also be used in other chromatin-related applications such as in vitro protein-DNA binding assays and nuclear enzyme assays.
- Extremely fast procedure – The entire procedure, from cell/tissue sample to ready-to-use chromatin, takes less than 60 minutes.
- Convenient and Flexible: The kit is suitable for preparing both native chromatin and cross-linked chromatin from cells or tissues in monolayer or suspension.
- The uncut chromatin makes it customizable for various analysis workflows that require intact or fragmented chromatin, including chip, in vitro protein-DNA interaction analysis, nuclear enzyme assay, etc.
Chromatin immunoprecipitation (ChIP) offers an advantageous tool for studying protein-DNA interaction. With ChIP, the experimenter can determine whether a specific protein binds to 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, measurement of the amount of histone H3 methylated in lysine 9 (meH3-K9) associated with a promoter region of a specific gene under various conditions can be achieved by a ChIP-PCR assay, while recruitment of meH3 -K9 to promoters on the ChIP-chip can detect a whole genome-scale. In particular, the ChIP method with specific antibodies directed against various transcription factors is in high demand.
Principle and procedure
The ChromaFlash ™ Chromatin Extraction Kit contains all the reagents necessary for successful chromatin extraction directly from mammalian cells or tissues. The sample cell membranes, with or without crosslinking, are decomposed using the provided lysis buffer. The 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.
Starting materials and input quantity
Starting materials can include various tissue or cell samples, such as flask or microplate culture cells, fresh and frozen tissues, etc. The number of cells and tissues for each preparation can be 1 x 105 to 5 x 106 cells and 10 mg to 200 mg, respectively. For optimal preparation, the input amount should be 1 to 5 x 10 6 cells or 50 to 200 mg of tissues. The chromatin yield is approximately 4 ug per 10 6 cells or per 50 mg of tissues.