DNA Demethylase Screening

DNA demethylation refers to the removal of a methyl group from DNA by DNA demethylases, including TDG (thymine DNA glycosylase),TET1 (methylcytosine dioxygenase 1) and TET2 (methylcytosine dioxygenase 2), which are pivotal in a range of DNA demethylation studies such as screening of inhibitors, enzyme kinetics, selectivity profiling, and protein arrays. The conversion of the modified base 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC) is the initial step of active demethylation of DNA. TET1 initiates this step and further oxidizes 5hmC to generate 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC), which can then both be converted backed to unmethylated cytosine. Global DNA demethylation exists in the developing zygote and primordial germ cells of the embryo, while some somatic cells undergo locus-specific DNA demethylation. DNA demethylation can be passive or active, and it is implicated with epigenetic reprogramming of genes and important disease mechanisms. 5-hmC levels are lower in several tumor tissues than normal tissues and its absence has been suggested as a possible biomarker for the diagnosis of cancer. Therefore, DNA demethylation studies could lead to important discoveries with implications for cancer therapy.

DNA demethylation pathway

Figure 1. DNA demethylation pathway (Wolffe, A. P.; et al. 1999)

Creative BioMart brings decades of experience in performing DNA demethylase assays to make your compounds advance speedily through drug development workflows. At Creative BioMart, our DNA demethylase screening service is based on the chemiluminescent technology with high quality and efficiency. Additionally, a large array of DNA demethylases and substrates produced in-house are provided to facilitate your epigenetic research.

Procedure of DNA Demethylases Screening at Creative BioMart

Highly specific antibodies that recognize demethylated products are involved in our demethylase assay. First, sample provided by clients is incubated with targeted enzyme and a biotinylated substrate. Acceptor beads, primary antibody and donor beads are then added sequentially. The following last step is to read the Alpha-counts.

General procedure of DNA demethylase screening assay

Figure 2. General procedure of DNA demethylase screening assay

Advantages of Our Service

  1. Screening for multiple inhibitors or targets by a panel of assays
  2. IC50 (half maximal inhibitory concentration) determination and single point concentration assay
  3. Large scale high throughput screening service
  4. Receive data after few days of compound submission
  5. Get project guidance in a time-efficient manner

As a world-class contract research organization in biopharmaceutical filed, Creative BioMart is an ideal partner to identify the optimal inhibitors for your target DNA demethylase and support your team to step towards critical milestone in epigenetic therapy projects. Professional and comprehensive screening service portfolios are delivered in a time-saving and cost-efficient manner. If you have any special requirements in DNA demethylase screening and profiling service, please don’t hesitate to contact us. We are looking forward to working together with your attractive projects.

1. Wolffe, A. P.; et al. DNA demethylation. PNAS. 1999, 96(11): 5894-5896.

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