Product Overview :
Methylated DNA Quantification Kit (Colorimetric) is used for measuring methylated DNA.
Description :
DNA methylation occurs by the covalent addition of a methyl group at the 5-carbon of the cytosine ring by DNA methyltransferases, resulting in 5-methylcytosine (5-mC). In somatic cells, 5-mC is found almost exclusively in the context of paired symmetrical methylation of the dinucleotide CpG, whereas in embryonic stem (ES) cells, a substantial amount of 5-mC is also observed in non-CpG contexts. The biological importance of 5-mC as a major epigenetic modification in phenotype and gene expression has been recognized widely. For example, decrease in 5-mC content (DNA hypomethylation) is likely caused by methyl-deficiency due to a variety of environmental influences, and has been proposed as a molecular marker in multiple biological processes such as cancer. It has been well demonstrated that the decrease in DNA methylation is one of the most important characteristics of cancer. Thus, the quantification of 5-mC content or methylation in cancer cells could provide very useful information for detection and analysis of this disease.
Applications :
The Methylated DNA Quantification Kit (Colorimetric) is suitable for detecting DNA methylation status using DNA isolated from any species such as mammalians, plants, fungi, bacteria, and viruses in a variety of forms including, but not limited to, cultured cells, fresh and frozen tissues, paraffin-embedded tissues, plasma/serum samples, and body fluid samples.
Usage :
For research use only (RUO)
Storage :
Upon receipt: (1) Store ME3, ME4, ME6, and ME7 at –20°C away from light; (2) Store ME1, ME5, ME8, and 8-Well Assay Strips at 4°C away from light; (3) Store remaining components (ME2 and ME9) at room temperature away from light.All components of the kit are stable for 6 months from the date of shipment, when stored properly.Note: Check ME1 (10X Wash Buffer) contains salt precipitates before use. If so, briefly warm at room temperature or 37°C and shake the buffer until the salts are re-dissolved.
Kit Components :
ME1 (10X Wash Buffer) 14 mlME2 (Binding Solution) 5 mlME3 (Negative Control, 20 μg/ml)* 10 μlME4 ( Positive Control, 20 μg/ml )* 10 μlME5 (Capture Antibody, 1000 μg/ml)* 4 μlME6 (Detection Antibody, 400 μg/ml)* 8 μlME7 (Enhancer Solution) 8 μlME8 (Developer Solution) 5 mlME9 (Stop Solution) 5 mlAssay Strip (with Frame) 48(8x6) wells* Spin the solution down to the bottom prior to use.Note: The ME3 Negative Control is an unmethylated polynucleotide containing 50% of cytosine. The ME4 Positive Control is a methylated polynucleotide containing 50% of 5-methylcytosine.
Detection method :
Colorimetric
Compatible Sample Types :
DNA Extract
Features & Benefits :
Colorimetric assay with easy-to-follow steps for convenience and speed. The entire procedure can be finished within 4 hours.Innovative kit composition enables background signals to be extremely low, which eliminates the need for plate blocking and allows the assay to be simple, accurate, reliable, and consistent.High sensitivity, of which the detection limit can be as low as 0.2 ng of methylated DNA (methylation).Optimized antibody and enhancer solutions allow high specificity to 5-mC, with no cross-reactivity to unmethylated cytosine and no or negligible cross-reactivity to hydroxymethylcytosine within the indicated concentration range of the sample DNA.Universal positive and negative controls are included, which are suitable for quantifying methylated DNA from any species.Strip-well microplate format makes the assay flexible: manual or high throughput analysis.
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