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EpiQuik m6A RNA Methylation Quantification Kit (Colorimetric)


For absorbance-based quantitation of N6-methyladenosine in RNA in an ELISA-like format

Citations (22) | (5) | Write a Review
Suggested Workflow
RNA Isolation
RNA Methylation Quantification
m6A standard control was added into the assay wells at different concentrations and then measured with the EpiQuik™ m6A RNA Methylation Quantification Kit (Colorimetric).
Schematic procedure of the EpiQuik™ m6A RNA Methylation Quantification Kit (Colorimetric).
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 EpiQuik™ m6A RNA Methylation Quantification Kit.
Input Type: RNA
Research Area: RNA Methylation
Target Application: Amount Quantitation
Vessel Format: 96-Well Plate
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-9005-4848 reactions $299.00 
P-9005-9696 reactions $499.00 
Availability: Usually Ships in 1 Day or Same day delivery Same Day NY Delivery 
Product Overview

The EpiQuik™ m6A RNA Methylation Quantification Kit (Colorimetric) is a complete set of optimized buffers and reagents to colorimetrically quantify N6-methyladenosine (m6A or 6mA) 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. The kit has the following advantages and features:

  • Colorimetric ELISA-like assay using an m6A antibody with easy-to-follow steps for convenience and speed. The entire procedure can be completed within 3 hours and 45 minutes.
  • High sensitivity, of which the detection limit can be as low as 10 pg of m6A.
  • Unique binding solution allows RNA >70 nts to be tightly bound to the wells, which enables quantification of m6A from both mRNA and ncRNA such as tRNA, rRNA, and snRNA.
  • 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.
  • Universal positive and negative controls are included, which are suitable for quantifying m6A from any species.
  • Strip-well microplate format makes the assay flexible for manual or high throughput analysis.
  • Simple, reliable, and consistent assay conditions.

Background Information 
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.

Principle & Procedure
In this assay, 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. Both negative and positive RNA controls are provided in this kit. 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. Because m6A content can vary from tissue to tissue, and from normal and diseased states, or vary under treated and untreated conditions, it is advised to run replicate samples to ensure that the signal generated is validated. This kit will allow the user to quantify an absolute amount of m6A and determine the relative m6A RNA methylation states of two different RNA samples.

Starting Materials
Starting materials can include various tissue or cell samples such as cells from flask or microplate cultured cells, fresh and frozen tissues, paraffin-embedded tissues, blood, body fluid samples, etc.

Product Components

WB (10X Wash Buffer)
BS (Binding Solution)
NC (Negative Control, 100 µg/ml)*
PC (Positive Control,  m6A 2 µg/ml)*
CA (Capture Antibody, 1000 X) *
DA (Detection Antibody, 1000 X)*
ES (Enhancer Solution)*
DS (Developer Solution)
SS (Stop Solution)
8-Well Assay Strips (With Frame)
User Guide

*Spin the solution down to the bottom prior to use.

User Guide & MSDS

[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

Product Citations

Guo J et. al. (June 2018). Deregulation of UBE2C-mediated autophagy repression aggravates NSCLC progression. Oncogenesis. 7(6):49.

Du J et. al. (June 2018). The regulation of skeletal muscle fiber-type composition by betaine is associated with NFATc1/MyoD. J Mol Med (Berl).

Dai T et. al. (May 2018). Analogous modified DNA probe and immune competition method-based electrochemical biosensor for RNA modification. Biosens Bioelectron. 114:72-77.

Faulds KJ et. al. (May 2018). Glycogen synthase kinase-3 (Gsk-3) activity regulates mRNA methylation in mouse embryonic stem cells. J Biol Chem.

Gu S et. al. (April 2018). N6-methyladenosine mediates the cellular proliferation and apoptosis via microRNAs in arsenite-transformed cells. Toxicol Lett.

Feng Z et. al. (March 2018). METTL3 regulates alternative splicing of MyD88 upon the lipopolysaccharide-induced inflammatory response in human dental pulp cells. J Cell Mol Med.

Ding C et. al. (January 2018). Increased N6-Methyladenosine causes infertility is associated with FTO expression. J Cell Physiol.

Zhou S et. al. (January 2018). FTO regulates the chemo-radiotherapy resistance of cervical squamous cell carcinoma (CSCC) by targeting β-catenin through mRNA demethylation. Mol Carcinog.

Lu N et. al. (January 2018). Curcumin Attenuates Lipopolysaccharide-Induced Hepatic Lipid Metabolism Disorder by Modification of m<sup>6</sup> A RNA Methylation in Piglets. Lipids. 53(1):53-63.

Lewinska A et. al. (August 2017). Reduced levels of methyltransferase DNMT2 sensitize human fibroblasts to oxidative stress and DNA damage that is accompanied by changes in proliferation-related miRNA expression. Redox Biol. 14:20-34.

Lewinska A et. al. (August 2017). Sulforaphane-Induced Cell Cycle Arrest and Senescence are accompanied by DNA Hypomethylation and Changes in microRNA Profile in Breast Cancer Cells. Theranostics. 7(14):3461-3477.

Haiyan Wang et. al. (July 2017). A sensitive photoelectrochemical immunoassay of N6 - methyladenosine based on dual-signal amplification strategy: Ru doped in SiO2 nanosphere and carboxylated g-C3N4 Biosensors and Bioelectronics.

Walters BJ et. al. (June 2017). The Role of The RNA Demethylase FTO (Fat Mass and Obesity-Associated) and mRNA Methylation in Hippocampal Memory Formation. Neuropsychopharmacology. 42(7):1502-1510.

Elkashef SM et. al. (May 2017). IDH Mutation, Competitive Inhibition of FTO, and RNA Methylation. Cancer Cell. 31(5):619-620.

Slobodin B et. al. (April 2017). Transcription Impacts the Efficiency of mRNA Translation via Co-transcriptional N6-adenosine Methylation. Cell. 169(2):326-337.e12.

Zhang S et. al. (March 2017). m6A Demethylase ALKBH5 Maintains Tumorigenicity of Glioblastoma Stem-like Cells by Sustaining FOXM1 Expression and Cell Proliferation Program Cancer Cell.

Lewinska A et. al. (February 2017). Downregulation of Methyltransferase Dnmt2 Results in Condition-dependent Telomere Shortening and Senescence or Apoptosis in Mouse Fibroblasts. J Cell Physiol.

Ishihara Y et. al. (November 2016). Hsa-miR-520d-5p promotes survival in human dermal fibroblasts exposed to a lethal dose of UV irradiation npj Aging and Mechanisms of Disease. (2)

Yin H et. al. (October 2016). Electrochemical immunosensor for N6-methyladenosine detection in human cell lines based on biotin-streptavidin system and silver-SiO2 signal amplification. Biosens Bioelectron.

Zhang C et. al. (October 2016). Hypoxia-inducible factors regulate pluripotency factor expression by ZNF217- and ALKBH5-mediated modulation of RNA methylation in breast cancer cells. Oncotarget. 7(40):64527-64542.

Zhang C et. al. (April 2016). Hypoxia induces the breast cancer stem cell phenotype by HIF-dependent and ALKBH5-mediated m6A-demethylation of NANOG mRNA. Proc Natl Acad Sci U S A. 113(14):E2047-56.

Yuen KC et. al. (January 2016). NIPBL Controls RNA Biogenesis to Prevent Activation of the Stress Kinase PKR. Cell Rep. 14(1):93-102.

Customer Reviews

Rating by c*******@gmail.com Unverified Purchase Reviewed on: Monday 05 March, 2018
Application Description
This is our go-to kit. The instructions are easy to follow, the results are reproducible, and it is a quality product of good value. We will definitely buy it again.
Rating by y*************@163.com Verified Purchase Reviewed on: Friday 19 January, 2018
Application Description
Use to detect the change of m6A levels after knockdown of METTL3 in cancer cells

Rating via Distributor/Reseller Verified Purchase Reviewed on: Monday 25 September, 2017
Application Description
Using this kit to measure the m6A content of RNA

This description is very clear, detailed ;step by step according to the protocol to do, very simple;

but I have a suggestion ,maybe you can highlight the key place with other colors marked to remind;

Other Thoughts
once the reagent is open ,it will expire in a several mouth,and you can’t realized it .you need pay attention!

The horizontal axis is the experimental group and the control group, and the vertical axis is m6A content
Rating by k**********@osu.edu Unverified Purchase Reviewed on: Tuesday 28 March, 2017
Application Description
Works as its supposed to. I tried it and its good. I will rebuy it.
Rating by c*********@ucsf.edu Verified Purchase Reviewed on: Thursday 02 February, 2017
Application Description
We used it to measure methylation level of mRNA. The user guide is user friendly, rated 5. The product presented great, rate 5. The overall rate of the product is 5.

The pros of the product: easy to use, the result is reproducible, instruction is easily to follow

Cons of this product: none, we just don't use it very often

Other Thoughts
The delivery was quick and the rep was very friendly and responded quickly when we had questions about ordering.
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