The Thermo Scientific Pierce Agarose ChIP Kit provides a complete set of reagents and a simple, fast and reproducible protocol to perform chromatin immunoprecipitation (ChIP) assays.
Features of the Agarose ChIP Kit:
• Simple and fast ChIP assay protocol (< 8 hours) • Highly efficient isolation and lysis of nuclei • Easy and reproducible enzymatic digestion • Low-background and high-binding capacity Protein A/G agarose resin • Highly specific positive controls included: RNA polymerase II antibody and GAPDH PCR primers • Fast and reproducible spin-column format • High-recovery DNA purification • ChIP-validated antibodies available
The Pierce Agarose ChIP Kit contains sufficient reagents to perform 30 ChIP assays with appropriate controls using an optimized protocol and a convenient spin-column format. ChIP-validated and quality-guaranteed antibodies are also available for use the the Pierce Agarose ChIP Kit. Because antibodies that work for western blotting may not function in ChIP assays, we have assembled our most popular ChIP-validated monoclonal and polyclonal antibodies here to help you move from experimental setup to results more quickly.
The strength of the ChIP assay is its ability to capture a snapshot of specific protein-DNA interactions as they occur in living cells, and then quantitate the interactions using standard or quantitive PCR. A successful ChIP assays requires a number of critical steps to be carried out (crosslinking, chromatin preparation, immunoprecipitation) prior to detecting the target genomic DNA sequence. Individually, each step in the ChIP assay protocol can be time consuming to optimize. The Pierce Agarose ChIP Assay Kit simplifies the process, making accurate and reproducible results easier to obtain.
To perform ChIP assays with the Pierce Agarose ChIP Kit, protein-DNA complexes are stabilized and then extracted. In vivo crosslinking is traditionally achieved with formaldehyde. Crosslinking performed directly in cells locks in the protein-DNA complexes, trapping these unstable and sometimes transient interactions. To lyse, extract and solubilize the crosslinked complexes, the ChIP assay kit includes the Thermo Scientific Chromatin Prep Module (also sold separately, Part No. 26158). Together, this complete ChIP assay kit provides a simple, reliable and convenient means for isolating chromatin-bound DNA and enriching samples for the proteins of interest with less than 15% contamination from other cellular compartments - without a Dounce homogenizer.
Sufficient For: 30 complete chromatin IP assay experiments• ChIP Grade Protein A/G Plus Agarose, 0.65 mL (store at 4°C) • IP Dilution/Wash Buffer (5X), 11 mL (store at 4°C) • IP Wash Buffer 3 (5X), 4.5 mL (store at 4°C) • IP Elution Buffer (2X), 4.5 mL (store at 4°C) • Column Accessory Pack, 3 packs (store at room temperature or 4°C) • Microcentrifuge Tubes (1.5mL), 75 tubes (store at room temperature or 4°C) • DNA Clean-Up Column, 40 columns (store at room temperature or 4°C) • DNA Column Binding Solution, 30 mL (store at room temperature or 4°C) • DNA Column Wash Solution, 6 mL (store at room temperature or 4°C) • pH Indicator, 0.8 mL (store at room temperature or 4°C) • DNA Column Elution Solution, 5 mL (store at room temperature or 4°C) • Anti-RNA Polymerase II Antibody, 25 µL (store at -20°C) • Normal Rabbit IgG (1 mg/mL), 10 µL (store at -20°C) • ChIP Positive Control Primers (GAPDH promoter, Human-specific), 100 µL (store at -20°C) • Pierce Chromatin Prep Module (Part No. 26158) (store Proteinase K and Micrococcal Nuclease at -20°C. Store all other components at 4°C).