title: Quantitative measurement of genome-wide DNA methylation by a reliable and cost-efficient enzyme-linked immunosorbent assay technique.
abstract: DNA methylation, the conversion of cytosine to 5-methylcytosine, is an important epigenetic modification involved in gene regulation. DNA methylation is essential for normal development whereas abnormal methylation has been implicated in pathological conditions including cancer. To evaluate the extent and variation of genome-wide DNA methylation and its changes during cellular differentiation and tumorgenesis as well as the interplay with histone modifications, accurate and reproducible quantification of the genomic DNA methylation level is required. These measurements have so far been achieved only by sophisticated and costly techniques. Here we report the generation of an enzyme-linked immunosorbent assay (methDNA-ELISA) for the accurate quantification of global DNA methylation levels. The linear region of this methDNA-ELISA ranges from 1 to 10%, making it highly suitable for the typical ranges from 2 to 6% in mammalian genomes. This method requires 10 ng of isolated DNA per sample, thus permitting investigation with minimal amounts of DNA previously not applicable for global DNA methylation analysis, e.g., clinical biopsies or cells collected by microdissection.
authors.affiliation: Institute for Molecular Medicine, Research Group Immunobiology, Duesseldorf, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Kremer
authors.forename: S
authors.initials: S
authors.lastname: Metzger
authors.forename: V
authors.initials: V
authors.lastname: Kolb-Bachofen
authors.forename: D
authors.initials: D
authors.lastname: Kremer
mesh_terms: D044503:5-Methylcytosine / Q000378:metabolism*; D045744:Cell Line, Tumor; D003362:Cost-Benefit Analysis; D003596:Cytosine / Q000378:metabolism; D004247:DNA / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000191:economics / Q000379:methods*; D005786:Gene Expression Regulation; D015894:Genome, Human; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA