EZ-Magna ChIP A/G Chromatin Immunoprecipitation Kit

General description

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.

Unlike standard ChIP protocols that can be laborious and time consuming, the Magna ChIP kit protocol can reduce the amount of time required to perform a ChIP experiment from three days to one. Additionally, the smaller Magna ChIP reaction volume increases the relative concentration of the antibody enabling the ChIP reaction to be performed with reduced amounts of both antibody and sheared chromatin. Finally because this kit uses a blend of protein A and protein G beads, a wider range of antibody isotypes can be used than A or G alone. This allows a wider variety of antibodies to be used and avoids the need to purchase separate kits for protein A and protein G based immunoprecipitation. Because Magna ChIP kits use paramagnetic beads they are compatible with automated high throughput platforms, thus allowing a large number of ChIP reactions to be carried out simultaneously. Features & Benefits:

• Magnetic bead-based rapid protocol allows performance of ChIP in as little as 1 day
• Blend of protein A+G bead blend allows ChIP using a broader range of antibodies than A or G alone
• Negative and positive control antibodies and control primer set to simplify validation of experimental procedure
• Includes spin columns to make DNA purification easier and more reliable - no more messy phenol-chloroform extractions
• Complete kit with all required reagents for reliable and reproducible results
• Compatible with ChIPAb+ validated antibody and primer sets

Chromatin Immunoprecipitation (ChIP) is an important technique allowing the analysis of 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.

Application

Research Category
Epigenetics & Nuclear Function

Used to detect/quantify: Protein A+G

Packaging

Kit capacity: 22 chromatin immunoprecipitation assays

Components

Magnetic Protein A/G Beads

ChIP Dilution Buffer

Low Salt Wash Buffer

High Salt Wash Buffer

LiCl Wash Buffer

TE Buffer

Cell Lysis Buffer

Nuclear Lysis Buffer

ChIP Elution Buffer (w/o Proteinase K)

Proteinase K

RNase A

10X Glycine

10X PBS

Protease Inhibitor Cocktail II

Anti-RNA polymerase II, clone CTD4H8

Normal Mouse IgG

Control Primers

Spin Filters

Collection Tubes

Bind Reagent A

Wash Reagent B

Elution Reagent C

Physical form

Two boxes containing 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).

Storage and Stability

Upon 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.

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless 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.