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Active Motif's DNA methylation assays offer fast, reliable solutions to streamline your methylation analysis. To see additional product details or data, please click on highlighted links below.
- MethylCollector™ is a unique technology that facilitates methylated DNA enrichment from a variety of cell and tissue types.
- MethylCollector™ Ultra is an enhanced version of our MethylCollector™ Kit that improves the enrichment of CpG-methylated DNA compared to current MBD or antibody-based methods.
- UnMethylCollector™ is a novel technology that allows for positive identification of unmethylated CpGs using sonicated or enzymatically digested genomic DNA.
- MethylDetector™ features our optimized bisulfite conversion protocol and a simplified DNA recovery method.
- DNMT Activity / Inhibition Assay to detect DNMT1, DNMT3a & DNMT3b DNA methyltransferase activity.
- Fully Methylated Jurkat DNA is used as a positive control in DNA methylation assays.
- DNA Methylation Antibodies – check out our growing list of antibodies related to DNA methylation.
DNA methylation is an epigenetic event
DNA methylation is an epigenetic event that affects cell function by altering gene expression. During methylation, methyl groups are transferred from the cofactor, S-adenosyl-L-methionine, to the fifth-carbon of cytosine in a CpG dinucleotide. This reaction is catalyzed by one of the DNA methyltransferase enzymes. Studying DNA methylation is of considerable importance since aberrant methylation is prevalent in many human cancers, and because methylation is also involved in embryonic development and cell cycle regulation.
CpG islands are small regions of the DNA in which the CpG dinucleotide frequency is higher than would normally be expected. While CpG islands are only found in approximately 1% of the genome, they coincide with more than 60% of human promoters. CpG islands are normally not methylated, however, if a CpG island within a promoter becomes methylated the gene associated with the promoter is permanently silenced, and this silencing can be transmitted through mitosis. This means that CpG island methylation is an epigenetic means of inheritance.
Due to the association of DNA methylation in development and disease, much research depends on the ability to accurately quantify DNA methylation. Active Motif offers several techniques that can be employed for DNA methylation analysis.
One such method is bisulfite conversion using Active Motif's MethylDetector™ Kit. This is a process in which unmethylated cytosines are converted into uracils, while methylated cytosines remain unchanged. The DNA is then amplified by PCR and analyzed by sequencing or restriction digest. A comparison of the sequences of converted and untreated DNA will reveal the methylation profile of the sample.
Another method is the isolation of methylated CpG islands using recombinant methyl-binding protein MBD2b, or the MBD2b/MBD3L1 complex, as in the MethylCollector™ product line. Genomic DNA is fragmented by either sonication or enzymatic digest and combined with the His-tagged methyl-binding protein in order to enrich for methylated CpG dinucleotides. The enriched DNA is suitable for use in many downstream applications, such as analysis using endpoint or realtime PCR.
Because CpG islands are normally unmethylated, the ability to enrich for unmethylated DNA fragments allows researchers to validate the methylation status of the locus of interest and identify hypomethylated DNA. While other techniques confirm methylation of CpG dinucleotides, a negative result is often designated as unmethylated DNA. With Active Motif's unique UnMethylCollector™ kit, unmethylated promoters can now be verified using a recombinant His-tagged CXXC protein, from the mouse Mbd1, which specifically binds unmethylated DNA. This DNA can be analyzed by either endpoint or realtime PCR, or used in a variety of other applications.
Active Motif also offers fully methylated Jurkat DNA that can be used as a positive control in methylation analysis studies. Additionally, Active Motif offers a growing list of DNA methylation antibodies.
