Note: Storage Temperature - see Storage/Stability section for individual component storage conditions
It has been well demonstrated that DNA methylation plays an important role in the regulation of gene expression. At any given time about 70% of all CpG dinucleotides in the mammalian genome are methylated.1 Hypo- and hyper-methylation across the genome contribute to changes in gene expression. Determining the methylation status of DNA is an important tool in epigenetic studies.8,11
Shifts away from normal global DNA methylation levels have been observed in various cancers, neurological disorders, autoimmune diseases (such as lupus and cardiovascular diseases), and aging.10
The Imprint Methylated DNA Quantification Kit provides a high-throughput, non-radioactive means of measuring global DNA methylation shifts in about half of a day.
This kit contains all reagents required to detect relative levels of methylated DNA. Up to 200 ng of purified DNA is bound to the wells of the assay strip. The methylated DNA is detected using the Capture and Detection antibodies, then quantified colorimetrically. The amount of methylated DNA present in the sample is proportional to the absorbance measured.
A methylated DNA control is provided. A standard curve can be performed (range: 10-100 ng) or a single quantity of DNA can be used as a positive control. Because global methylation can vary from tissue to tissue, and from normal and diseased states, it is advised to run replicate samples at different DNA concentrations to ensure that the signal generated falls within the detection range of the plate reader used.
The kit will allow the user to determine relative methylation states of two different DNA samples.
1 x Wash Buffer
For the 48-reaction kit, add 11 mL of 10x Wash Buffer to 99 mL of Water, Molecular Biology Reagent. Store at room temperature for up to six months.
For the 96-reaction kit, add 22 mL of 10x Wash Buffer to 198 mL of Water, Molecular Biology Reagent. Store at room temperature for up to six months.
Dilute sample or control DNA in DNA Binding Solution. Each well can bind a maximum of 200 ng of total DNA.
Methylated DNA Capture
To calculate methylation levels relative to the Methylated Control DNA, use the following steps.
Single Point Method
Percent methylation relative to the Methylated Control DNA
Standard Curve Method
ng Methylated DNA in sample(s) from standard curve of Methylated Control DNA
Imprint is a registered trademark of Sigma-Aldrich Co. LLC.
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