Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: DNA
Product namem6A DNA Methylation Assay Kit (Colorimetric)
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.
Storage instructionsPlease refer to protocols.
Components 1 x 48 tests 1 x 96 tests 10X Wash Buffer 1 x 14ml 1 x 28ml 8-Well Assay Strips (With Frame) 6 units 12 units Binding Solution 1 x 5ml 1 x 10ml Capture Antibody, 1000 X 1 x 5µl 1 x 10µl Detection Antibody, 1000 X 1 x 6µl 1 x 12µl Developer Solution 1 x 5ml 1 x 10ml Enhancer Solution 1 x 5µl 1 x 10µl Negative DNA Control, 100 µg/ml 1 x 10µl 1 x 20µl Positive Control, 200 µg/ml containing m6A 1 µg/ml 1 x 10µl 1 x 20µl Stop Solution 1 x 5ml 1 x 10ml
ab233488 has not yet been referenced specifically in any publications.