Description
Key facts
Detection method:Fluorescent,
Sample types:Tissue, Suspension cells, Other biological fluids, Adherent cells,
Assay type:Quantitative,
Sensitivity:= 5 pg/well,
Assay time:3h 45m,
Assay Platform:Microplate reader
Product details:
m6A RNA Methylation Assay Kit (Fluorometric) (ab233491) is a complete set of optimized buffers and reagents to fluorometrically quantify methylated N6-methyladenosine (m6A) in RNA. It is suitable for a direct detection of m6A RNA methylation status using total RNA isolated from any species such as mammals, plants, fungi, bacteria and viruses.
This kit contains a unique binding solution allowing RNA >70 nts to be tightly bound to the wells, which enables quantification of m
A from both mRNA and nc-RNA such as tRNA, rRNA and snRNA. The optimized antibody and enhancer solutions allow high specificity to m
A, with no cross-reactivity to unmethylated adenosine within the indicated concentration range of the sample RNA. Also included are universal positive and negative controls which are suitable for quantifying m
A from any species.
N6-methyladenosine (m
A) is the most common and abundant modification in RNA molecules present in eukaryotes. The m
A modification is catalyzed by a methyltransferase complex METTL3 and removed by the recently discovered m
A RNA demethylases FTO and ALKBH5, which catalyze m
A demethylation in an α-ketoglutarate (α-KG)- and Fe2+-dependent manner. It was shown that METTL3, FTO, and ALKBH5 play important roles in many biological processes, ranging from development and metabolism to fertility. m
A accounts for more than 80% of all RNA base methylations and exists in various species. m
A is mainly distributed in mRNA and also occurs in non-coding RNA such as tRNA, rRNA, and snRNA. The relative abundance of m
A in mRNA transcripts has been shown to affect RNA metabolism processes such as splicing, nuclear export, translation ability and stability, and RNA transcription. Abnormal m
A methylation levels induced by defects in m
A RNA methylase and demethylase could lead to dysfunction of RNA and diseases. For example, abnormally low levels of m
A in target mRNAs due to increased FTO activity in patients with FTO mutations, through an as-yet undefined pathway, contributes to the onset of obesity and related diseases. The dynamic and reversible chemical m6A modification in RNA may also serve as a novel epigenetic marker of profound biological significance. Therefore, more useful information for a better understanding of m
A RNA methylation levels and distribution on RNA transcripts could benefit diagnostics and therapeutics of disease.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
N6-methyladenosine (m6A) is a chemical modification present in RNA molecules specifically marking the adenosine base with a methyl group at the nitrogen-6 position. This modification weighing approximately 14 Da occurs in diverse eukaryotic species and influences RNA metabolism. m6A methylation can be detected using techniques like m6A dots blot or m6A ELISA. The modification is abundant in tissues like the brain and testis reflecting its critical role in varying cell types. Also m6A is known as a dynamic and reversible mark with its levels continuously adjusted by enzymes called writers (methyltransferases) erasers (demethylases) and readers (RNA-binding proteins).
Biological function summary
M6A methylation affects mRNA processing stability translation and decay. It integrates into large multi-protein complexes where it influences gene expression outcomes by affecting the RNA's interaction with the cellular machinery. This methylation modification acts as a regulatory signal that influences essential processes such as cell differentiation and circadian rhythms. Elucidating the biological functions of m6A involves studying how it affects RNA fate and its downstream gene regulatory networks.
Pathways
M6A modification is central to the mRNA metabolic pathway and the PI3K-Akt signaling pathway. It interacts with various proteins such as METTL3 an m6A methyltransferase which is vital for mediating m6A modification. It also interacts with YTH domain-containing proteins that recognize m6A marks influencing transcript dynamics and gene expression. The interplay of m6A with proteins in these pathways underlines its role in fine-tuning cellular processes and responses.
M6A modification has a significant impact on cancer and neurological disorders. In cancers alterations in m6A methylation patterns can promote oncogenic transformation and metastasis. m6A-related proteins such as FTO an m6A demethylase have shown connections to these pathways affecting cancer progression. In neurological disorders m6A impacts aspects of neural development and function and abnormalities in its regulation may contribute to diseases like Alzheimer's. Understanding the roles of m6A in diverse diseases can pave the way for novel therapeutic approaches.
