Overview

  • Product name

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

    Fluorescent
  • Sample type

    Other biological fluids, Tissue, Adherent cells, Suspension cells
  • Assay type

    Quantitative
  • Sensitivity

    5 pg/well
  • Assay time

    3h 45m
  • Product overview

    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 m6A from both mRNA and nc-RNA such as tRNA, rRNA and snRNA. The optimized antibody and enhancer solutions allow high specificity to m6A, 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 m6A from any species.

  • 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

  • Storage instructions

    Please 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
    Detector Antibody, 1000 X 1 x 6µl 1 x 12µl
    Dilution Buffer 1 x 4ml 1 x 8ml
    Enhancer Solution 1 x 5µl 1 x 10µl
    Fluoro Developer 1 x 8µl 1 x 16µl
    Fluoro Enhancer 1 x 8µl 1 x 16µl
    Negative Control, 100 µg/mL 1 x 10µl 1 x 20µl
    Positive Control, m6A 2 µg/mL 1 x 10µl 1 x 20µl
  • Research areas

  • Relevance

    N6-Methyladenosine (m6A) is an abundant modification found in mRNA, tRNA, snRNA, as well as long non-coding RNA, in all species. RNA adenosine methylation is catalyzed by a multicomponent complex composed of METTL3/MT-A70, METTL14, and WTAP in mammals. METTL3 & METTL14 are responsible for the methyltransferase activity of the complex, and WTAP mediates substrate recruitment.
  • Alternative names

    • m6A
    • N6-methyladenosine

Images

Protocols

References

ab233491 has not yet been referenced specifically in any publications.

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