Description
Key facts
Detection method:Colorimetric,
Assay type:Quantitative,
Sensitivity:= 0.01 ng,
Range:10 - 500 ng/mL,
Assay time:3h 45m,
Assay Platform:Microplate reader
Product details:
m6A ELISA Kit / m6A RNA Methylation Assay Kit ab185912 is suitable for the direct detection of m6A RNA methylation status using total RNA isolated from any species such as mammals, plants, fungi, bacteria, and viruses.
Be aware that m6A is present in mRNA and ribosomal RNA, and this kit detects m6A in total RNA, ie. both mRNA and ribosomal RNA and any other RNA containing m6A.
The amount of RNA for each assay can be 100 ng - 300 ng. For optimal quantification, the input RNA amount should be 200 ng, as the abundance of m6A is generally less than 0.1% of total RNA.
m6A ELISA / m6A RNA methylation assay protocol summary
- Total RNA is bound to strip wells using a RNA high binding solution
- N6-methyladenosine (m6A) is detected using a specific capture N6-methyladenosine antibody and detection antibody
- The detected signal is enhanced and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer at a wavelength of 450 nm. The amount of m6A is proportional to the OD intensity measured.
- A standard curve can be performed (range: 0.02 to 1 ng of m6A) or a single quantity of m6A can be used as a positive control.
How other researchers are using
m6A RNA Methylation Quantification Kit (Colorimetric) has been used in a variety of sample type including:
- Tumor and control peritoneal tissues 1
- Human osteosarcoma cancer cell lines 143B and U2OS 2
- Human lung epithelial cell line BEAS-2B, Human bronchial epithelial cell line 16HBE 3
References:
1 - Reamon-Buettner S et al. 2024; 2 - Mei Z et al. 2024; 3 - Ma L et al. 2024.
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.
