m6A (N6-methyladenosine) ELISA Kit

N6-methyladenosine (m6A) is the most prevalent internal modification of messenger RNAs (mRNAs), affecting their fold, stability, degradation, and cellular interaction(s). m6A RNA plays an essential role in both physiological and pathological conditions, especially in the initiation and progression of different types of human cancers. In recent years, m6A in DNA has been defined as another important epigenetic and epitranscriptomic marker in higher eukaryotes. A significant decrease in m6A levels has also been reported in a variety of human cancer cells. Increasing evidence shows that dynamic m6A modification in genomic DNA is associated with brain functions, glioblastoma, embryogenesis, reproduction, and embryonic stem cell development, and cardiovascular disease. Thus, determining the global changes in m6A levels is very necessary to fully understand the physiological and pathological status of biology, such as human disease.

The RayBio^® m6A ELISA Kit is a competitive enzyme immunoassay developed for rapid detection and quantitation of m6A in urine, serum, or plasma samples collected from humans, rats, or mice. The quantity of m6A is determined by a known m6A standard curve. The m6A kit has a high sensitivity of approximately 0.435 ng/ml. The linearity, stability, precision and spiking & recovery were also confirmed and showed good performance. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown samples.

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Kit Components

• Microplate reader capable of measuring absorbance at 450 nm
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1-liter graduated cylinders
• Absorbent paper
• Distilled or deionized water
• SigmaPlot software (or other software which can perform four-parameter logistic regression models)
• Tubes to prepare standard or sample dilutions
• Orbital shaker
• Aluminum foil
• Plastic wrap

1. Prepare all reagents, samples and standards as instructed.
2. Add 50 μL standards or samples to appropriate well. Incubate for 10 minutes at room temperature.
3. Add 50 μL anti-m6A to appropriate well. Incubate 1 hour at room temperature. And wash 5 times.
4. Add 100 μL prepared HRP-Secondary antibody solution. Incubate 1 hour at room temperature. And wash 5 times.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 2-10 minutes at room temperature.
6. Add 100 μL Stop Solution to each well. Read at 450 nm immediately.

The entire kit may be stored at -20°C to -80°C for up to 6 months from the date of shipment. For extended storage, it is recommended to store it at -80°C. Avoid repeated freeze-thaw cycles.