Overview

  • Product name

    m6A RNA Methylation Assay Kit (Colorimetric)
    See all N6-methyladenosine (m6A) kits
  • Detection method

    Colorimetric
  • Assay type

    Quantitative
  • Sensitivity

    0.01 ng
  • Range

    0.01 ng/ml - 0.5 ng/ml
  • Assay time

    3h 45m
  • Product overview

    The m6A RNA Methylation Quantification Kit (Colorimetric) (ab185912) is a complete set of optimized buffers and reagents to colorimetrically 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.


    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 tha 0.1% of total RNA.

  • Notes

    N6-methyladenosine (m6A) is the most common and abundant modification in RNA molecules present in eukaryotes. The m6A modification is catalyzed by a methyltransferase complex METTL3 and removed by the recently discovered m6A RNA demethylases FTO and ALKBH5, which catalyze m6A 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. m6A accounts for more than 80% of all RNA base methylations and exists in various species. m6A is mainly distributed in mRNA and also occurs in non-coding RNA such as tRNA, rRNA, and snRNA. The relative abundance of m6A in mRNA transcripts has been shown to affect RNA metabolism processes such as splicing, nuclear export, translation ability and stability, and RNA transcription. Abnormal m6A methylation levels induced by defects in m6A RNA methylase and demethylase could lead to dysfunction of RNA and diseases. For example, abnormally low levels of m6A 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 m6A RNA methylation levels and distribution on RNA transcripts could benefit diagnostics and therapeutics of disease.

  • Platform

    Microplate reader

Properties

Images

  • Quantification of m6A RNA methylation in different samples. 200 ng of RNA isolated from different tissues or cells were added into the assay wells and the m6A contained in RNA was measured using the m6A RNA Methylation Quantification Kit (Colorimetric).

Protocols

References

This product has been referenced in:

  • Dorn LE  et al. The N6-Methyladenosine mRNA Methylase METTL3 Controls Cardiac Homeostasis and Hypertrophy. Circulation 139:533-545 (2019). Read more (PubMed: 30586742) »
  • Kmietczyk V  et al. m6A-mRNA methylation regulates cardiac gene expression and cellular growth. Life Sci Alliance 2:N/A (2019). Read more (PubMed: 30967445) »
See all 4 Publications for this product

Customer reviews and Q&As

Answer

The ab185912 m6A RNA Methylation Quantification Kit (Colorimetric) has been optimized for testing m6A RNA Methylation from total RNA. It has not been tested specifically with purified mRNA.

It is important to note that total RNA consists of 80% ribosomal RNA and about 3% mRNA. So you may need much larger samples in order to obtain the 100- 300 ug required for the assay.

I am sorry we have no testing data from mRNA to help answer your question in more detail in this case. 

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