The chromatin immunoprecipitation (ChIP) assay is a powerful method for analyzing epigenetic modifications and genomic DNA sequences bound to specific regulatory proteins. Briefly, in ChIP protein-DNA complexes are crosslinked, immunoprecipitated, purified, and amplified for gene- and promoter-specific analysis of known targets using real time PCR or sequencing or for the identification of new target sequences. In a related microarray-based method, ChIP-on-chip, the purified immunoprecipitated DNA is labeled and hybridized to a variety of high resolution microarray types.

A chemical fixation step with UltraPure™ reagents is used to covalently crosslink a transcription factor to a promoter sequence, followed by sonication/fragmentation of DNA for bead-based immunoprecipitation.

For best results, chromatin should be sheared to uniform 100-250 bp fragments. 500-1kb for qPCr 200-250 for sequencing.

Immunopreciptiation - Simple and Efficient Isolation of Antibodies and their Target Proteins

Dynabeads® provide simple robust, and rapid small-scale purification of immunoglobulins and subsequent isolation of target protein. These superparamagnetic, polystyrene beads offer gentle magnetic handling with minimal physical stress on the target proteins, and are a key component of the MAGnify™ Chromatin Immunoprecipitation System.

ChIP-qualified Antibodies
In addition, Invitrogen offers a number of additional primary antibodies directed against DNA-binding proteins that have not yet  been tested in Chromatin Immunoprecipitation. Search for antibodies.

Reverse Crosslink and Purify DNA - Using Simple and Reliable Reagents
Experience improved results using Invitrogen’s Proteinase K and reverse crosslinking steps.

Gene-specific Analysis

After the immunoprecipitation step, different PCR-based approaches can be used in gene-specific analysis to determine how much DNA of interest has been precipitated. Real-time PCR amplification is frequently a preferred technique. The EXPRESS SYBR® GreenER™ system is specially formulated to offer the best sensitivity, specificity, and reproducibility for your real-time qPCR analysis

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