m6A DNA Methylation Assay Kit (Colorimetric)
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(2 Publications)
m6A DNA Methylation Assay Kit (Colorimetric)(ab233488) kit contains all reagents necessary for the quantification of m6A in DNA.
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KIAA1752, FTO, Alpha-ketoglutarate-dependent dioxygenase FTO, Fat mass and obesity-associated protein, U6 small nuclear RNA (2'-O-methyladenosine-N(6)-)-demethylase FTO, U6 small nuclear RNA N(6)-methyladenosine-demethylase FTO, mRNA (2'-O-methyladenosine-N(6)-)-demethylase FTO, mRNA N(6)-methyladenosine demethylase FTO, tRNA N1-methyl adenine demethylase FTO, m6A(m)-demethylase FTO
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Functional Studies - m6A DNA Methylation Assay Kit (Colorimetric) (AB233488)
Example of a m6A standard curve.
m6A standard control was added into the assay wells at different concentrations and then measured with the m6A DNA Methylation ELISA Kit (Colorimetric) (ab233488).
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Functional Studies - m6A DNA Methylation Assay Kit (Colorimetric) (AB233488)
Quantification of m 6A content of various human DNA samples with the m6A DNA Methylation ELISA Kit (Colorimetric) (ab233488).
Product details
m6A DNA Methylation Assay Kit (Colorimetric)(ab233488) kit contains all reagents necessary for the quantification of m6A in DNA. In this assay, DNA is bound to strip wells using DNA high binding solution. m6A is detected using capture and detection antibodies. The detected signal is enhanced and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer. The amount of m6A is proportional to the OD intensity measured.
N 6 -methyladenosine (m6A) is the most common and abundant modification on RNA molecules present in eukaryotes. DNA m 6A is also identified in multicellular eukaryotes including Caenorhabditis elegans and Drosophila melanogaster, and furthermore identified in higher eukaryotes including plants, mouse and human cells. m6A plays crucial roles in regulating DNA replication, transposition, transcription, and cellular defense. In humans, the DNA m 6A modification is most likely catalyzed by a methyltransferase complex METTL3 and removed by the α-ketoglutarate (α-KG)- and Fe2+ -dependent dioxygenases such as ALKBH5 and TET-like enzymes. It was shown that METTL3 and α-KG /Fe2+ - dependent dioxygenases play important roles in many biological processes, ranging from development and metabolism to fertility.
The dynamic and reversible chemical m6A modification on DNA may also serve as a novel epigenetic marker of profound biological significance. Down-regulation of m 6A modification was first characterized in human cancer cells and tissues, relative to their normal controls. m 6A is found to be the most regulated DNA modification in cancers. In addition to the regulation in cancer cells, relative to the primary cell/tissues which contain quite low amounts of DNA m 6A (<0.001%), a hundreds-fold increase of m 6A modification was found for in vitro cultured human cells (0.03%-0.22%). Therefore, identifying m 6A DNA methylation levels and distribution on DNA could advance understanding of epigenetic regulation of biological process at the genomic level, and further provide useful information for improving diagnostics and therapeutics of disease.
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Biological function summary
The m6A modification of DNA affects transcriptional activity by influencing the interaction between DNA and proteins. This type of methylation acts similarly to a switch turning gene expression on or off as needed. In several cases m6A works closely within complexes involving RNA-binding proteins and transcription factors coordinating the stability splicing and translation of mRNAs. This suggests a significant role in post-transcriptional gene regulation. For instance components such as METTL3 and METTL14 are associated with m6A methylation and contribute to forming a complex that performs these regulatory functions.
Pathways
The modification by m6A DNA methylation appears to be an integral part of epigenetic pathways that govern cellular differentiation and development. It also influences the methylation signaling pathway aiding in the fine-tuning of epigenetic states within the cell. Proteins related to m6A-modified pathways include ALKBH5 and FTO both of which demethylate m6A marks allowing dynamic control of DNA interpretation. Understanding these modifications within pathways requires considering how they affect gene regulatory networks on an epigenetic level.
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Publications (2)
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Nanomaterials (Basel, Switzerland) 11: PubMed34835707
2021
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International journal of molecular sciences 22: PubMed33670900
2021
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