m1A (N1-methyladenosine) ELISA Kit

RNA modifications affect RNA metabolism and are involved in many different biological processes. Mono-methylation of adenosine in the N1 position, N1-methyladensoine (m1A), is a reversible modification that was first discovered in tRNA in 1966. Decades later, m1A was found in tRNA, rRNA, mitochondrial RNA and mRNA. It has been verified that m1A dysregulation affects multiple cellular processes, including RNA structural stability, folding, interactions with proteins, cell viability, impaired self-renewal ability, cell proliferation, and cell death. They tend to be conserved and have been implicated in many critical biological processes including development, disease, circadian rhythm and embryonic stem cell fate transition, as well as age-related disorders such as Alzheimer’s disease.

Today, it is recognized that the dynamic regulation of m1A in response to physiological stress and the dysregulated expression of m1A regulators are correlated with tumorigenesis and cancer recurrence. Urine samples of various cancers, rheumatoid arthritis, and AIDS patients have all demonstrated high levels of m1A, which supports its role as a functional detection biomarker. The urinary m1A correlates with m1A level in tissue from the same patients with esophageal cancer. Furthermore, elevated serum levels of m1A have been detected under stress conditions. Recent m1A research is uncovering new roles and mechanisms involving m1A. However, the complete role of 1-methyladenosine as a modified nucleoside has yet to be elucidated.

The RayBio® m1A ELISA Kit is a competitive enzyme immunoassay developed for rapid detection and quantitation of m1A in urine, serum, or plasma samples collected from human, rat or mouse. The quantity of m1A is determined by a known m1A standard curve. The m1A kit has a high sensitivity of approximately 2.544 ng/ml. The performance was also verified to exhibit good characteristics in terms of linearity, stability, precision, and spiking & recovery. 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-m1A 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. For prepared reagent storage, see table below.