Chromatin Immunoprecipitation (ChIP) is a standard technique for analysis of transcriptional regulation and chromatin structure.
Chromatin Immunoprecipitation (ChIP) is a standard technique for analysis of transcriptional regulation and chromatin structure. All standard ChIP methods utilize agarose-based affinity beads to capture the antibody/chromatin complexes. However, spin-washing these beads is slow and tedious. In addition, DNA isolated through these methods must be further purified (by spin columns or phenol/chloroform extraction & precipitation) prior to PCR analysis.
We have developed a greatly simplified ChIP protocol that utilizes magnetic protein G-coated beads for antibody capture. This dramatically reduces washing time and enables the use of multi-channel pipetting, so that many samples can be assayed simultaneously (Figure 1).
Reduced background eliminates steps
The new ChIP-IT™ Express method improves on traditional ChIP by reducing or eliminating several time-consuming steps. The magnetic beads have much lower background than traditional agarose beads, so pre-clearing and blocking of the chromatin are no longer necessary. Wash steps are faster and fewer because centrifugation is replaced by magnetic pull-down, significantly reducing the amount of hands-on time. In addition, improvement of the chromatin elution and proteinase K treatment steps has completely eliminated the need for a separate DNA purification step after ChIP is complete (Figure 2). This not only saves time, it minimizes manipulations and eliminates DNA loss that occurs during purification, both of which help ensure sample-to-sample consistency.
Complete kit ensures successful ChIP
ChIP-IT Express Kits transform ChIP from a labor-intensive protocol that is appropriate for only a small number of samples into a rapid method that is suitable for experiments requiring multiple samples (e.g. studies of pathway induction, time-course experiments, etc.). The kits incorporate all of these improvements and include the reagents required to perform your choice of either enzymatic- or sonication-type chromatin shearing, and perform 25 ChIP reactions, including a powerful bar magnetic. For complete details, visit Web site (external).
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