Background: Epigenetic changes refer to stable, heritable, and reversible modifications. DNA methylation is one such epigenetic change and has been shown to be associated with almost every biological process. DNA methylation can increase the functional complexity of prokaryotic and eukaryotic genomes by providing additional avenues for the control of cellular processes. DNA methylation is also dynamic and thus can control the timing of cellular events. In prokaryotes, DNA methylation occurs at the C-5 or N-4 positions of cytosine, as well as the N-6 position of adenine. Multiple enzymes that are capable of methylating bacterial DNA have been identified and are named DNA-MTases. Initially, DNA methylation in bacteria was reported to be associated with restriction-modification systems, wherein foreign DNA that lacks DNA methylation can be recognized and degraded by host methylation-sensitive restriction enzymes. Further studies in bacteria also implicated DNA methylation in the maintenance of the DNA replication fidelity, gene expression regulation, and virulence. In plants, DNA methylation is found in the sequence context of CpG or CpNpG and has been implicated in normal plant development and regulation of transcription and transposition. At least two classes of DNA methyltransferases (DNMTs) have been identified and characterized in plants. These include the MET1 family of methyltransferases, which preferentially methylate cytosine in CpG context and function both as de novo and maintenance DNMTs, and a second class of DNMTs, named chromomethylases (CMTs), which are unique to plants and methylate cytosine in a CpNpG sequence context. In mammalian cells, DNA methylation is predominantly found at CpG dinucleotides, most of which reside within gene promoter regions, but in some instances, such as in the case of mouse and human embryonic stem cells, DNA methylation can also be found in non-CpG contexts. DNA methylation in mammalian cells is associated with repression of transcription and maintenance of genomic stability. Due to its important role in genome maintenance and other biological processes, deregulation of DNA methylation is associated with multiple human diseases including cancer. Three major enzymes are required for either de novo methylation (DNMT3A and DNMT3B) or maintenance methylation (DNMT1) in mammalian cells. There are several methods of detecting total 5-methylcytosine content in the genome. DNA can be digested into single nucleotides and total genomic 5-methylcytosine can be quantitated by high performance liquid chromatography, thin-layer chromatography, or liquid chromatography/mass spectroscopy, but all of these methods are labor intensive and technically demanding. Global DNA Methylation ELISA Kit is a competitive enzyme immunoassay developed for rapid detection and quantitation of 5'-methyl-2'-deoxycytidine (5MedCyd) in urine directly, or extracted cell or tissue DNA samples. The quantity of 5MedCyd in an unknown sample is determined by comparing its absorbance with that of a known 5MedCyd standard curve. The kit has a 5MedCyd detection sensitivity range of 78 nM to 20,000 nM. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown samples.

Principle of the Assay: The Global DNA Methylation ELISA Kit is a competitive ELISA for the quantitative measurement of 5MedCyd. The unknown 5MedCyd samples or 5MedCyd standards are first added to the 5MedCyd DNA Conjugate Coated plate. After a brief incubation, an Anti-5MedCyd antibody is added, followed by an HRP conjugated secondary antibody. The 5MedCyd content in unknown samples is determined by comparison with a predetermined 5MedCyd standard curve.

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Small volumes of Global DNA Methylation elisa kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS168205 is a ready-to-use microwell, strip-or-full plate ELISA (enzyme-linked immunosorbent assay) Kit for analyzing the presence of the Global DNA Methylation, ELISA Kit target analytes in biological samples. The concentration gradients of the kit standards or positive controls render a theoretical kit detection range in biological research samples containing Global DNA Methylation. The ELISA analytical biochemical technique of the MBS168205 kit is based on Global DNA Methylation antibody-Global DNA Methylation antigen interactions (immunosorbency) and an HRP colorimetric detection system to detect Global DNA Methylation antigen targets in samples. The ELISA Kit is designed to detect native, not recombinant, Global DNA Methylation. Appropriate sample types may include undiluted body fluids and/or tissue homogenates, secretions. Quality control assays assessing reproducibility identified the intra-assay CV (%) and inter-assay CV(%).