Chromatin Immunoprecipitation (ChIP)
Gain further insight into the dynamic interactions between transcription proteins and components of chromatin, and ultimately understand their roles in cellular functions
One of the most widely used methods for studying chromatin-related modifications is chromatin immunoprecipitation (ChIP), a process by which a protein of interest binds to a specific genomic DNA region. This popular ChIP assay provides epigenetic researchers with valuable information about the interaction among certain proteins or protein modifications and a specific DNA sequence.
Protein-DNA interactions are crucial to certain cellular functions such as DNA replication and recombination, gene transcription, chromosome segregation, signal transduction, and epigenetic silencing. Understanding protein-DNA interactions can be used, for example, to compare the amount of histone methylation linked to a specific gene promoter region among two different groups, such as tissues that are diseased or healthy. Chromatin immunoprecipitation methods can also be used to map the genomic location of various other histone modifications.
Chromatin is a combination of DNA and proteins that packages DNA into a smaller volume to fit in the cell, helps to prevent DNA damage, and controls gene expression and DNA replication. Proper isolation of chromatin samples is an important factor in assays for studying protein-DNA interactions.
Chromatin immunoprecipitation (ChIP) is an antibody-based method used for determining the location of DNA binding sites on the genome for a particular protein of interest. This technique is a convenient means for studying protein-DNA interactions that occur inside the nucleus of cells and for understanding cellular processes. Downstream applications of ChIP include ChIP-sequencing, ChIP-PCR, and ChIP-on-chip (microarrays).
Measurement of direct interactions between protein and DNA in vitro has an advantage in analyzing the binding of different transcription factors to specific DNA consensus sequences located in the gene promoters. By investigating protein-DNA interaction in vitro, it is possible to identify the genetic targets of DNA, which leads to a better understanding of cellular processes.
Chromatin immunoprecipitation is a powerful technique for studying protein-DNA interaction in vivo, and can be coupled with various downstream applications to profile and map histone methylation patterns.
Chromatin immunoprecipitation can be used as a tool to identify activated genes associated with acetylated histones, and can be coupled with qualitative and quantitative PCR, MSP, DNA sequencing, and Southern blot as well as DNA microarrays to further profile or map histone acetylation patterns.
Methyl-CpG-binding domain protein 2 (MBD2) is a member of the MBD protein family and has been shown to catalyze demethylation by direclty removing methyl groups from 5-methylcytosine residues in DNA. Chromatin immunoprecipitation would be useful in the identification of silenced genes associated with MBD2, and can be combined with qualitative and quantitative PCR, MSP, DNA sequencing, DNA microarrays, and Southern blot to profile or map MBD2 binding patterns.
The accessibility of regulatory elements in chromatin is critical for many aspects of gene regulation. Nucleosomes positioned over regulatory elements inhibit access of transcription factors to DNA. To elucidate the role of the interactions between chromatin and transcription factors, it is crucial to determine chromatin accessibility through mapping of the nucleosome positioning along the genome.
Apply SNAP-ChIP DNA-barcoded recombinant designer nucleosomes (dNucs) as next-generation spike-ins for chromatin immunoprecipitation (ChIP).
Apply best-in-class SNAP-ChIP Certified Antibodies with exquisite target specificity, less than 20% cross-reactivity and more than 5% IP efficiency.