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ChIP-IT™ Kits make performing chromatin immunoprecipitation (ChIP, chromatin IP) more successful by combining all of the critical components needed in a single kit, including shearing components. However, depending on how many immunoprecipitations you perform with each sample of sheared chromatin, you may run out of shearing components before using up all of the immunoprecipitation components in the ChIP-IT Kits. For this reason, the ChIP-IT Sonication & Enzymatic Shearing Kits are also sold separately. This enables you to buy only the reagents you need, while giving you the flexibility to use either traditional sonication or our unique enzymatic shearing.
| Name | Format | Cat. No. | Price | |
|---|---|---|---|---|
| ChIP-IT™ Shearing Kit for sonication (included in 53001, 53004 & 53008) | 10 rxns | 53002 | $180 |
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| Enzymatic Shearing Kit (included in 53006, 53007 & 53009) | 10 rxns | 53005 | $205 |
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Choose from sonication or reproducible enzymatic shearing
The first step in successful ChIP is shearing the chromatin into 200-1000 bp fragments. This has traditionally been performed by subjecting the isolated chromatin to different pulses of sonication. Although sonication is an effective method for shearing DNA, it can be difficult to optimize due to complications arising from emulsification and overheating. And, because the quality of your sheared sample depends greatly upon the quality of your sonicator, it may be necessary to purchase an expensive, "high-end" sonicator to get reproducible shearing. Because of this, Active Motif has developed a more robust and user-friendly method to shear chromatin for ChIP. The Enzymatic Shearing Kit uses a proprietary Enzymatic Shearing Cocktail that quickly shears DNA into 200-1000 bp fragments (Figure 1). Because enzymatic shearing is solely time and temperature dependent, the problems associated with sonication are eliminated and ChIP results are improved.
Figure 1: Analysis of DNA sheared using the Enzymatic Shearing Kit.
HeLa cells were fixed for 10 minutes with 1% formaldehyde and chromatin was prepared using the Enzymatic Shearing Kit. Chromatin was sheared with the Enzymatic Shearing Cocktail for 5, 10 & 15 minutes. The sheared and unsheared chromatin samples were subjected to cross-link reversal, treated with Proteinase K, phenol/chloroform extracted and precipitated as described in the protocol. Samples were separated by electrophoresis through a 1% agarose gel to assess shearing results:
Lane 1: 100 to 1000 bp ladder.
Lane 2: Unsheared HeLa DNA.
Lane 3: HeLa DNA treated for 5 minutes.
Lane 4: HeLa DNA treated for 10 minutes.
Lane 5: HeLa DNA treated for 15 minutes.
Flexible ordering options
The ChIP-IT Sonication and Enzymatic Shearing Kits provide sufficient shearing components to optimize shearing conditions and then make 5 preparations of sheared chromatin from three 15 cm plates (4.5 x 107 cells); each chromatin preparation will yield enough material to perform approximately 16 ChIP reactions. (These identical shearing components are already included in the ChIP-IT and ChIP-IT Express Kits, so you don’t need to buy a stand-alone shearing kit unless you’re running out of shearing components.)
Contents & Storage
Shearing Buffer (or Digestion Buffer and Enzymatic Shearing Cocktail), 10X PBS, 10X Glycine, 1X Lysis Buffer, Protease Inhibitor Cocktail, PMSF, EDTA, Proteinase K and RNase A. Reagent storage conditions vary from room temperature to -20°C. Please download a manual for complete kit specifications and storage conditions. All reagents are guaranteed stable for 6 months when stored properly.
