SpeedRun One-Hour m6A RNA ELISA Kit (Colorimetric)

The SpeedRun™ One-Hour m6A RNA ELISA Kit (Colorimetric) is a complete set of optimized reagents designed for detecting and quantifying N6-methyladenosine (m6A) RNA methylation directly from total RNA isolated from any species, including mammals, plants, fungi, bacteria, and viruses. The kit can be used with mRNA as well as non-coding RNA, including tRNA, rRNA, and snRNA. The kit has the following advantages and features:

• Fast: Colorimetric assay with easy-to-follow steps for convenience and speed. The entire procedure can be completed in 1 hour*
• Sensitive: High sensitivity, of which the detection limit can be as low as 10 pg of m6A
• Versatile: Unique binding solution allows that RNA >70 nts can be tightly bound to the wells, which enables quantification of m6A from both mRNA and nc-RNA such as tRNA, rRNA and snRNA
• Specific: 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
• Convenient: Universal positive and negative controls are included, which are suitable for quantifying m6A from any species
• Flexible: Strip-well microplate format makes the assay flexible for manual or high throughput analysis
• Reliable: Simple, reliable, and consistent assay conditions

Background Information
N6-methyladenosine (m6A) is the most common and abundant RNA modification in eukaryotes. m6A methylation is catalyzed by the methyltransferase complex METTL3 and can be removed by the m6A RNA demethylases FTO and ALKBH5. m6A accounts for more than 80% of RNA base methylations and is found in a variety of RNA species. It is mainly distributed in mRNA but 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 important RNA metabolism processes, including splicing, nuclear export, translation, stability, and transcription. Abnormal m6A methylation levels resulting from altered RNA methylase or demethylase activity can lead to RNA dysfunction and have been associated with disease. To address the need for accurate and convenient m6A measurement, EpigenTek offers a series of m6A RNA quantification kits and further refines its RNA methylation assay expertise with the development of the SpeedRun™ One-Hour m6A RNA ELISA Kit (Colorimetric).

Principle & Procedure
The kit contains all reagents necessary for the quantification of m6A in RNA. In this assay, total RNA is bound to strip wells using high binding solution. m6A is detected using the m6A Detection Complex. The detected signal is enhanced and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer. The amount of m6A is proportional to the OD intensity measured.

Starting Materials
Input material can be total RNA isolated from any species. The amount of RNA for each assay can range from 100 to 300 ng per reaction, with 200 ng recommended for optimal quantification.