		Transcriptional Regulation Co-Immunoprecipitation DNA Repair Luciferase Reporter Assay Nuclear Receptor ELISAs Supershift & Gelshift Assays Transcription Factor DNA-binding ELISAs Epigenetics & Chromatin Chromatin Assembly Chromatin IP DNA Damage – Phospho-H2AX DNA Methylation Histone Acetyltransferase Activity (HAT) Histone Deacetylase Activity (HDAC) Histone Purification Protein Modification GTPase Activation Assay Kinase Assay Substrate Mitosis Assays Nitric Oxide Quantitation Phospho-Protein Detection Sandwich ELISAs SUMOylation Antibodies, Extracts & Proteins Antibodies Cell Extracts Recombinant Proteins Fluorescence & Labeling Albumin Quantification Cell Viability Assay Fluorescent Dyes & Labeling Immunofluorescence Live Cell Membrane Staining Mitosis Determination Protein and Extract Quantification Protein Labeling & Translocation Sample Preparation Fluorescent Dye Labeling Mitochondrial Separation mRNA Isolation Normalization Nuclear & Cytoplasmic Extraction Protein & Extract Quantification Transfection & Expression Competent E. coli Gene Silencing Protein Transfection Staining Reagents Bioinformatics & Text Mining TimeLogic® Biocomputing Solutions AKS 2

ChIP-IT™ Express Kits

magnetic beads make ChIP fast, easy & more reproducible

ChIP-IT™ Express Kits use protein G-coated magnetic beads instead of traditional agarose beads, making it possible to perform ChIP in just 1 day. The result is that ChIP-IT Express makes ChIP more successful while greatly reducing your time and effort.

Faster, more reproducible protocol
Choice of enzymatic or sonication methods for shearing of chromatin
Variety of control ChIP kits available for your model organism
Ready to ChIP chromatin available

ChIP-IT Express improves ChIP

ChIP-IT Express improves on traditional ChIP by reducing or eliminating several time-consuming steps. The kit's magnetic beads have much lower background than traditional agarose beads, so pre-clearing and blocking are no longer necessary. Washing is easier because the spin steps have been replaced by rapid magnetic pull-down. And, only 3 washes are required, as compared to 8 washes with traditional ChIP. This dramatically reduces the amount of hands-on time during the assay. This not only saves time, but it minimizes manipulations and eliminates the DNA loss that can occur during purification, helping to ensure sample-to-sample consistency.

Optimized buffers reduce background

While the magnetic beads and bar magnet included in ChIP-IT Express make ChIP much faster and easier, many of the kit's buffers have been optimized to provide you with better ChIP results. The Elution and Reverse Cross-linking Buffers help reduce the background, which enables you to discern effects that are smaller in magnitude. Reduced background also makes it possible to work with fewer cells (see Figure 2 in OVERVIEW).

In addition to reduced background, ChIP-IT Express buffers eliminate the need for DNA purification, so you can simultaneously elute the DNA from the beads while reversing the cross-links. These simple protocol changes give you results quickly and easily while offering the freedom to simultaneously perform multiple ChIP assays: now you can even perform multiple ChIPs in PCR tubes with a multi-channel pipettor (Figures 1 & 2).

Figures 1 & 2: Multiple-sample ChIP using ChIP-IT Express.

Immunoprecipitation and wash steps are fast and easy with magnetic beads because the pellet forms on the side of the tube, even pulling completely out of the buffer. Because it is simple to remove the buffer without disturbing the pellet, it is now possible to perform multiple ChIPs in PCR strips using a multi-channel pipettor.

Powerful magnet

ChIP-IT Express Kits include a strong bar magnet that can be used to turn pipette tip boxes into magnetic stands for immunoprecipitation and washing. P1000 tip boxes can be used with Eppendorf tubes (Figure 3), while P200 boxes work with PCR strips (Figures 1 & 2). Commercially available magnetic tube stands, such as those from Ambion (Figure 4) and Promega, can also be used.

Figure 3: Use of a standard tip box for magnetic ChIP.

ChIP-IT Express Kits include a strong bar magnet that can be used to convert pipette tip boxes into a magnetic stand for washing the protein G-coated magnetic beads.

Figure 4: Use of an Ambion 6-tube Magnetic Stand with ChIP-IT Express.

Commercially available magnetic tube racks, such as those from Ambion and Promega, can be used with ChIP-IT Express.

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 using sonication. Although sonication is effective, many users find that complications caused by emulsification and overheating can lead to variable results. To improve reproducibility, Active Motif has developed a unique enzymatic shearing method. For your convenience, all ChIP-IT and ChIP-IT Enzymatic Kits are available in your choice of sonication or enzymatic shearing formats and stand-alone kits are also available. Click here to learn more about your options. And, to make you ChIP even simpler, we also offer Ready-to-ChIP Chromatin. Using our chromatin and ChIP-IT reagents, the only variable in your ChIP will be the antibody you are testing.

ChIP-IT Control Kits and ChIP-validated antibodies

The original ChIP-IT Kits contain human positive and negative controls; these simplify the task of troubleshooting your experiment, interpreting your results and validating antibodies for use in ChIP. However, once an antibody has been shown to work in ChIP, it may no longer be necessary to run the controls. Moreover, many of our customers have requested ChIP controls for species other than human. For these reasons, ChIP-IT Express Kits are provided without controls. By purchasing a species-specific ChIP-IT Control Kit separately you can get exactly what you want, without having to pay for controls that you may not need. In addition, Active Motif offers an ever-increasing number of ChIP-validated antibodies, which are ideal for use with all of our ChIP-IT Kits.

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