Thermo Scientific Pierce Magnetic ChIP Kit provides a method to efficiently isolate chromatin-bound DNA via immunoprecipitation (chromatin IP). Convenient method for subsequent quantitative analysis by PCR.

Features of the Magnetic ChIP Kit:

•Fast—Purified DNA suitable for quantitative PCR can be obtained in about 8 hours
•Efficient and reproducible—Micrococcal nuclease digestion and nuclear lysis processes are highly optimized
•Sensitive—only 10,000 cells (1 x 10^4 ) to get results
•Low background—Pierce Protein A/G magnetic beads are blocked in DNA-free reagents to minimize background
•Comprehensive—Includes optimized positive control reagents: RNA Polymerase II Antibody and GAPDH Promoter PCR Primers

Pierce Magnetic Chromatin Immunoprecipitation Assay (ChIP) Kit contains sufficient reagents to be used The optimized protocol performs 30 ChIP assays using appropriate controls. The kit contains reagents for lysing cells, capturing protein-DNA complexes, reversing cross-links, and isolating DNA. Blocked Pierce Protein A/G Magnetic Beads have extremely high binding capacity and low non-specific background and can be used with a wide range of antibodies. These beads can be used in manual workflows with magnetic stands or automated platforms (e.g., Thermo Scientific KingFisher instruments). Quality-assured antibodies validated for ChIP are also available for use with Pierce Magnetic ChIP Kits.

Includes:
Kit contains reagents for chromatin preparation, IP and DNA purification, as well as positive controls (antibodies and primers)

Requires:
In vivo cross-linking agent (e.g. formaldehyde), microtip sonicator (e.g. Misonix™ Ultrasonicator 3000), preferred ChIP compliant antibodies, PCR primers for the target DNA sequence, PCR master mix (containing a dye such as SYBR™ Green if qPCR is required), and qPCR instrument

Application:
• By PCR or ChIP-Seq to identify sites of specific protein-DNA interactions on genomic DNA
• Monitor the effects of histone modifications or chemicals on DNA binding

To successfully perform ChIP assays, you need to Several critical steps (cross-linking, chromatin preparation, immunoprecipitation) are performed before targeting genomic DNA. Individually, each step in a ChIP protocol is time-consuming to optimize. Thermo Scientific Pierce Magnetic ChIP Kits simplify the ChIP process for accurate and reproducible results.

Protein-DNA complexes are first cross-linked in vivo with formaldehyde. The kit contains reagents that lyse cells and extract and solubilize cross-linked complexes. The complexes are then incubated with specific antibodies and finally separated using Pierce Protein A/G magnetic beads. After reversing cross-linking and digesting the protein, the resulting DNA fragments are purified and ready for standard or quantitative PCR.

More product data
•Androgen-dependent and -independent regulation of transcriptional activity Analysis

Related Products
Pierce™ Magnetic ChIP Kit
Pierce™ ChIP Grade Protein A/G Magnetic Beads