m7G (N7-methylguanosine) ELISA Kit

Currently, more than 170 modifications have been identified on RNA, which affect RNA metabolism and are involved in many different biological processes. As one of the prevalent posttranscriptional modifications of RNA, N7-methylguanosine (m7G) plays essential roles in RNA processing, metabolism, and function. m7G is the most ubiquitous mRNA cap modification and is also present in internal mRNA, microRNA (miRNA), tRNA and rRNA. m7G is positively charged and produced by the methyltransferase-like 1 (METTL1) and WD repeat domain 4 (WDR4) complex. Emerging evidence suggests that the METTL1/WDR4 complex promoted or inhibited the processes of brain development and diseases, as well as many kinds of tumor, including head and neck, lung, liver, colon, bladder cancer, and teratoma. Additionally, m7G is also crucial for tumor chemoresistance, vasculogenesis and prognosis. Therefore, m7G modification can be used as biomarkers or potential intervention targets, providing new possibilities for early diagnosis and treatment of tumors.

The RayBio® m7G ELISA Kit is a competitive enzyme immunoassay developed for rapid detection and quantitation of m7G in urine, serum, plasma, and RNA samples collected from human, rat or mouse. The quantity of m7G is determined by a known m7G standard curve. The m7G kit has a high sensitivity of approximately 0.164 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-m7G 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.