Urine m6A Assay Kit (ab233493)
Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 4 hr
- Sample type: Urine
- Sensitivity: 0.01 ng/well
Overview
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Product name
Urine m6A Assay Kit -
Detection method
Colorimetric -
Sample type
Urine -
Assay type
Quantitative -
Sensitivity
0.01 ng/well -
Assay time
4h 00m -
Species reactivity
Reacts with: Human -
Product overview
Urine m6A Assay Kit (ab233493) is a complete set of optimized buffers and reagents to colorimetrically quantify m6A in urine using an inhibitory competitive immunoassay method. It is suitable for detecting total urinary m6A levels, resulting from whole body turnover or degradation of DNA/RNA containing m6A, using urine from humans and animals.
In this ELISA-like inhibitory competitive immunoassay, urine samples and the m6A standard are first incubated with a m6A antibody solution and then transferred to strip wells coated with m6A polynucleotide. The well is washed to remove any unbound reagents after incubation and then a detection antibody is added to generate a signal that can be measured colorimetrically by reading the absorbance in a microplate spectrophotometer. Because m6A in the urine sample inhibits the binding of m6A antibody to m6A coated on the well, higher concentrations of m6A in the urine sample lead to a reduced binding of the antibody to the m6A on the well. Therefore the signal or OD intensity measured from the well will be inversely proportional to the amount of m6A in the urine sample and the amount of m6A in the urine sample can be quantified by a comparison with a predetermined m6A standard.
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Notes
Nucleobase m6A, a modified form of adenosine converted by adenosine methyltransferases is widespread in different cellular RNAs and also found in DNA. The biological importance of DNA/RNA m6A-methylation as a major epigenetic modification in phenotype and gene expression has been recognized widely. m6A plays crucial roles in regulating DNA replication, DNA damage, RNA splicing, transposition, transcription, and cellular defense. In humans, the m6A modification is probably catalyzed by a methyltransferase complex METTL3/METTL14 and removed by the α-ketoglutarate (αKG)- and Fe2+-dependent dioxygenases such as FTO, ALKBH5 and TET-like enzymes. It was shown that METTL3 and α-KG /Fe2+-dependent dioxygenases play important roles in many biological processes, ranging from development and metabolism to fertility.
Urinary excretion of m6A is an indication of the whole body turnover or the degradation of DNA/RNA, especially tRNA. The urinary m6A level can be changed with a change of the bodies’ turnover of m6A DNA/RNA or alteration of cellular DNA/RNA m6A status. A number of studies have indicated that m6A excreted in urine has the potential to act as a cancer biomarker. For example, an elevated level of urinary m6A was observed in colorectal cancer patients with active disease states. -
Platform
Microplate reader
Properties
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Storage instructions
Please refer to protocols. -
Components 1 x 48 tests 1 x 96 tests 10X Wash Buffer 1 x 14ml 1 x 28ml Binding Solution 1 x 5ml 1 x 10ml Capture Antibody, 1000X 1 x 4µl 1 x 8µl Detection Antibody, 2000X 1 x 4µl 1 x 8µl Developer Solution 1 x 5ml 1 x 10ml Enhancer Solution 1 x 4µl 1 x 8µl m6A Assay Solution, 500X 1 x 10µl 1 x 20µl m6A Standard, 2 µg/ml 1 x 10µl 1 x 20µl Negative Control, 50 µg/ml 1 x 10µl 1 x 20µl Plate 1 (Assay Plate, flat bottom) 1 unit 1 unit Plate 2 (Sample Preparation Plate, round bottom) 1 unit 1 unit Stop Solution 1 x 5ml 1 x 10ml
Images
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m6A standard was added into the assay wells at different concentrations and then measured with Urine m6A Assay Kit (ab233493).
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m6A level is quantified from different volumes of human urine using Urine m6A Assay Kit (ab233493).