General descriptionChromatin Immunoprecipitation (ChIP) is an important technique allowing the researcher to analyze in vivo interactions of proteins with genomic DNA. Any chromatin-associated or DNA binding protein can be analyzed with this technique, provided a good antibody to the protein exists. One can measure different proteins localized to a specific region of the genome, or the genome wide distribution of a specific protein. Another powerful application of this technique is to analyze changes in histone modifications that correlate with processes like transcription, mitosis or DNA repair.
Chromatin Immunoprecipitation (ChIP) is a powerful technique for mapping the in vivo distribution of proteins associated with chromosomal DNA. These proteins can be histone subunits and post-translational modifications or other chromatin associated proteins such as transcription factors, chromatin regulators, etc. Additionally, ChIP can be used to identify regions of the genome associated with these proteins, or conversely, to identify proteins associated with a particular region of the genome. ChIP methodology often involves protein-DNA and protein-protein cross-linking, fragmentation of the cross-linked chromatin, and subsequent immunoprecipitation of chromatin with an antibody specific to a target protein. The DNA fragments isolated in complex with the target protein can be identified by a variety of methods including PCR, DNA microarray and DNA sequencing. Standard or quantitative PCR can be performed to verify whether a particular DNA sequence (the gene or region of the genome) is associated with the protein of interest. The combination of ChIP and promoter or genomic tiling microarrays (ChIP-chip) allows genome-wide identification of DNA-binding sites for chromatin-associated proteins with precise resolution. Alternatively, high-throughput sequencing of libraries constructed from immunoprecipitated chromosomal DNA (ChIP-Seq) is a powerful alternative to ChIP-chip in mapping the protein-DNA interactions across mammalian genomes.
Features and Benefits- Easier: Spin columns make DNA purification easier and more reliable - no more messy phenol-chloroform extractions.
- Quicker: All reagents to process your samples are included - you don′t have to spend valuable time making them.
- Greater Reproducibility: Positive and negative control antibodies and PCR primers are included to help validate your results and to troubleshoot your experiments.
PackagingContains all necessary reagents to perform 22 individual chromatin immunoprecipitation (ChIP) reactions. Supplied buffers are sufficient to generate chromatin from up to five 15 cm plates of cultured cells, each plate providing up to 10 chromatin preparations (varies with cell and assay type).This EZChIP kit contains all necessary reagents to perform 22 individual chromatin immunoprecipitation (ChIP) reactions using inexpensive protein G agarose beads. Control primers included.
Kit capacity: 22 chromatin immunoprecipitation assays
Preparation NoteUpon receipt, store components at the temperatures indicated on the labels. Kit components are stable for 1 year from date of shipment when stored as directed.
Other Notes- DNA Purification Spin Columns and Collection Tubes
- ChIP Blocked Protein G Agarose
- Anti-RNA Polymerase II
- Control PCR Primers
- Normal Mouse IgG
- Bind, Wash and Elution Reagents
- Protease Inhibitor Cocktail II
- RNase A
- Proteinase K
- All required buffers
- ChIP Dilution Buffer
- Low Salt Immune Complex Wash Buffer
- High Salt Immune Complex Wash Buffer
- LiCl Immune Complex Wash Buffer
- TE Buffer
- 0.5M EDTA
- 5M NaCl
- SDS Lysis Buffer
- 1M Tris-HCl, pH 6.5
- 10X PBS
- 10X Glycine
- 1M NaHCO3
- Control Primers
- 20% SDS
- Spin Filters
- Collection Tubes
- Bind Reagent A
- Wash Reagent B
- Elution Reagent C
DisclaimerUnless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
