to BioTechniques free email alert service to receive content updates.

protocol
New Magnetic Bead—based Kits Expand Your Capabilities and Improve Your Results
Sponsored,vendor-submitted protocol   Published in November 2009 (p.25) DOI: 10.2144/000113003 Abstract

Chromatin immunoprecipitation (ChIP) is an important technique for the analysis of chromatin-associated proteins in their native chromatin context. Existing ChIP methods utilize agarose beads to capture the antibody/chromatin complexes, but spin-washing of agarose beads is tedious and slow. In addition, DNA isolated through these methods must be further purified (by laborious phenol/chloroform extraction and precipitation or using expensive spin columns) 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 multichannel pipetting, so that many samples can be assayed simultaneously (Figure 1).

Chromatin immunoprecipitation (ChIP) is an important technique for the analysis of chromatin-associated proteins in their native chromatin context. Existing ChIP methods utilize agarose beads to capture the antibody/chromatin complexes, but spin-washing of agarose beads is tedious and slow. In addition, DNA isolated through these methods must be further purified (by laborious phenol/chloroform extraction and precipitation or using expensive spin columns) 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 multichannel pipetting, so that many samples can be assayed simultaneously (Figure 1).

Reduced Background Eliminates Wash Steps

The 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 also minimizes manipulations and eliminates DNA loss that occurs during purification, both of which help ensure sample-to-sample consistency and the reproducibility of your experiments.





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.). This novel magnetic bead–based protocol is the basis for a family of groundbreaking kits that dramatically improve your ability to ask important questions about chromatin protein localization, transcription factor function, and the role of histone modifications in the regulation of genome function.

New ChIP Kits Expand Your Ability to Ask Important Questions

We have recently introduced two innovative new kits that enable you to ask questions that were previously difficult to address due to the technically challenging nature of the experiments. ChIP-IT Express HT allows you to rapidly and efficiently process a large number of ChIP reactions in a fast, reproducible high-throughput format. It combines the efficiency of the ground-breaking magnetic bead–based ChIP-IT Express Kit with a 96-well plate format. The new Re-ChIP-IT Kit enables sequential chromatin immunoprecipitations to be performed using two different antibodies, so you can assay for the simultaneous presence of two proteins or distinct histone modifications at the same genomic region of interest. These new kits are part of Active Motif's continuing commitment to develop cutting edge kits to perform your important experiments. For details on chromatin IP kits from Active Motif, visit www.activemotif.com/chipit.

Correspondence

Address correspondence to Active Motif, Inc., 1914 Palomar Oaks Way, Suite 150, Carlsbad, CA 92008, USA; Tel.: (877) 222-9543; [email protected], www.activemotif.com.

More Information
 
Do you have question about this protocol? Would you like more information about this protocol?
Submit your questions here and Biotechniques will forward your question and contact information from our database to an expert at
Yes, please forward my question, along with my name, title, company, and e-mail address to
BioTechniques will forward your question to the company, who will contact you directly by email.
Request more information
 
Name Question
Job Title
Company
Email
Protocol Categories
rightcontent.jsp called otherwise condition rightcontent.jsp called 222222222222222222222
Search Protocols
Search our protocols for your specific technique.
Submit Your Protocols
BioTechniques is always looking for author-submitted protocols related to peer-reviewed papers, so submit your protocols today to our editorial team. Suppliers or advertisers interested in publishing their protocols should also submit.
Click here for more information