ChIP-IT & ChIP-IT Express

faster, more successful chromatin immunoprecipitation

ChIP-IT Kits and reagents are a complete, flexible set of tools that make chromatin immunoprecipitation (ChIP) faster and more consistent. ChIP-IT Kits combine all of the critical components needed in a single kit, validated and proven to work in ChIP. Kits are available in your choice of sonication or our exclusive enzymatic shearing formats, either with or without controls.

In addition to the original ChIP-IT Kits, we offer ChIP-IT Express Kits, which utilize protein G-coated magnetic beads to greatly simplify and streamline the ChIP protocol. Because magnetic beads make it possible to reduce in length or completely eliminate a number of time-consuming steps, with ChIP-IT Express you can complete your ChIP in just 1 day. And, the recent addition of new elution buffers greatly reduces background, so you can now work with fewer cells (see Figure 2 below). And, to make ChIP even simpler, Active Motif also offers species-specific ChIP-IT Control Kits and Ready-to-ChIP Chromatin.

Click on a link below to go to specific information about the various ChIP-IT products, or keep reading for more general information on ChIP and ChIP-IT:

The ChIP-IT method

Chromatin immunoprecipitation involves the immunoprecipitation of protein/DNA complexes that have been stabilized via cross-linking. It offers a versatile solution by combining the specificity of immunoprecipitation, the sensitivity of PCR and the screening power of array profiling. However, ChIP can be technically challenging and difficult to validate without well-proven reagents. By providing proven antibodies, reagents and controls, ChIP-IT guarantees the best results possible.

In ChIP, intact cells are fixed using formaldehyde, which cross-links and therefore preserves protein/DNA interactions. DNA is then sheared into small uniform fragments and the DNA/protein complexes are immunoprecipitated using an antibody directed against the DNA-binding protein of interest. Following immunoprecipitation, the DNA is washed, cross-linking is reversed, and the proteins are removed by Proteinase K treatment. In the original ChIP-IT Kits, the DNA is then rapidly cleaned up using the included DNA purification columns. This DNA purification step in not necessary with ChIP-IT Express (Figure 1). The DNA is then screened to determine which genes were bound by the protein of interest.

Why use ChIP-IT?

• Complete solution all reagents and controls supplied

• Direct measurement of protein/DNA interactions, not just histone modifications

• Compatible with genome-wide profiling or selective PCR-based approaches

• No need to optimize reagents and protocol

• No phenol/chloroform extractions DNA purification columns included in original ChIP-IT Kits, DNA purification step eliminated in ChIP-IT Express

The ChIP-IT advantage

Classically, ChIP has been performed with antibodies directed against abundant chromatin components, such as acetylated histones. While this yields information about transcriptional activity of promoters, it does not reveal which transcription factor is bound to the promoter(s) of interest. In contrast, ChIP using transcription factor-specific antibodies enables direct monitoring of transcription factor/DNA interactions. Unfortunately, this is technically more challenging than classical ChIP, and requires the preparation of several complicated buffers, inhibitor cocktails and blocking reagents. In addition, result validation is difficult without antibodies, controls and a protocol proven to work in ChIP. To overcome these problems and enable ChIP of both transcription factors and histones, Active Motif's ChIP-IT Kits offer a complete solution by providing nearly everything needed to successfully perform ChIP experiments, including positive and negative controls (Figure 2).

Click on one of the links below to go to specific information about the various types of ChIP-IT Kits and reagents: