The gene-specific analysis techniques cannot provide information on the global level of DNA hydroxymethylation in the genome. Although the classical liquid chromatography tandem mass spectrometry/mass spectrometry (LC-MS/MS)-based approach to quantifying global DNA hydroxymethylation has high accuracy and repeatability but requires large numbers of DNA samples and currently not suitable for high-throughput analysis. Here, Creative BioMart is ready to provide global DNA 5hmC ELISA quantification service which is free of cross-reactivity with unmethylated cytosine or 5mC with short turnaround time, no matter what kind of samples the customers would like to detect.

The ELISA-based assay detects and quantifies 5hmC levels in genomic DNA from different sample types. This method is only suitable for the rough estimation of DNA hydroxymethylation. But it enables higher throughput processing of samples with only a small amount of genomic DNA per well.

What Is Global DNA 5hmC ELISA Quantification?

In this assay, genomic DNA is fragmented, denatured and bound to 96-well plates treated with special reagents to have a high DNA affinity. The primary antibody specific for 5hmC and the secondary antibody for tracking are sequentially added to detect the 5hmC fragments. The absorbance of each sample is read by a microplate spectrophotometer. The hydroxymethylation level of DNA samples can be determined by making a standard curve with DNA fragments containing known levels of 5hmC.

Features of Our Global DNA 5hmC ELISA Quantification

1. Tolerance of multiple sample types: we accept a wide range of biological and clinical samples, including but not limited to, genomic DNA isolated from a variety of species (mammals, plants, fungi, bacteria, and viruses, etc), cultured cells, fresh/frozen tissues, formalin-fixed and paraffin-embedded samples, plasma/serum samples, and body fluid samples
2. Specificity: high specificity to 5hmC, no cross-reactivity with unmethylated cytosine or methylated cytosine within the specified concentration range of sample DNA.
3. High sensitivity: this assay allows extremely low DNA sample input. The detection limit can be as low as 0.01% hydroxymethylated DNA of 100% input DNA.
4. Flexible: the microwell plate format allows the analysis to be used for manual or high-throughput analysis with higher throughput quantification than mass spectrometry.
5. Cost-effective: quantitative analysis by ELISA offers cost and speed advantages. It is a cheaper, more convenient alternative than mass-spectrometry or chromatography-based techniques.

Workflow of Global DNA 5hmC ELISA Quantification at Creative BioMart

Figure 1. Workflow of Global DNA 5hmC ELISA Quantification at Creative BioMart

With professional experts and strict quality control, global DNA 5hmC ELISA quantification service provided by Creative BioMart is ready for any customer demands with optimal customization. If you have any questions about this service, please feel free to contact us.