Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
Product nameGlobal RNA Methylation Assay Kit (5 Methyl Cytosine, Fluorometric)
Global RNA Methylation Assay Kit (5 Methyl Cytosine, Fluorometric) (ab233492) contains all reagents necessary for the quantification of global 5-mC RNA methylation. In the assay, RNA is bound to strip-wells that are specifically treated to have a high nucleic acid affinity. 5-mC in RNA is detected using capture and detection antibodies and then quantified fluorometrically by reading the fluorescence in a microplate spectrophotometer. The percentage of 5-mC RNA is proportional to the fluorescence intensity measured.
5-methylcytosine (5-mC) in DNA occurs by the covalent addition of a methyl group at the 5-carbon of the cytosine ring by DNA methyltransferases. This process has been well studied and is generally associated with repression of gene expression. It was also observed that in humans, 5-mC occurs in various RNA molecules including tRNAs, rRNAs, mRNAs and non-coding RNAs (ncRNAs). 5-mC seems to be enriched in some classes of ncRNA, but relatively depleted in mRNAs. Levels of 5-mC are variable in animal genomes, ranging from undetectable amounts in some insects to about 0.1-0.45% of total RNA in human cells. The majority (83%) of 5-mC sites were found in mRNAs. Within these transcripts 5-mC appears to be depleted within protein coding sequences but enriched in 5’ and 3’ UTRs. Two different methyltransferases, NSUN2 and Dnmt2 are known to catalyze 5-mC modification in eukaryotic RNA. There has been strong evidence that RNA cytosine methylation affects the regulation of various biological processes such as RNA stability and mRNA translation. Furthermore, loss of 5-mC in vault RNAs causes aberrant processing into Argonaute-associated small RNA fragments that can function as microRNAs. Thus, impaired processing of vault ncRNA may contribute to the etiology of human disorders related to NSun2-defciency.
Storage instructionsPlease refer to protocols.
Components 1 x 48 tests 1 x 96 tests 10X Wash Buffer 1 x 14ml 1 x 28ml 5-mC Antibody, 1000X 1 x 5µl 1 x 10µl 8-Well Assay Strips (With Frame) 1 x 6 units 1 x 12 units Binding Solution 1 x 5ml 1 x 10ml Dilution buffer 1 x 5ml 1 x 10ml Enhancer Solution, 1000X 1 x 5µl 1 x 10µl Fluoro Developer 1 x 12µl 1 x 24µl Fluoro Enhancer 1 x 12µl 1 x 24µl Negative Control containing 0% 5-mC, 100 µg/ml 1 x 50µl 1 x 100µl Positive Control containing 2% 5-mC, 100 µg/ml 1 x 10µl 1 x 20µl Signal Indicator, 1000X 1 x 5µl 1 x 10µl
ab233492 has not yet been referenced specifically in any publications.