MethylFlash Global DNA Methylation (5-mC) ELISA Easy Kit (Colorimetric)

Fast, reliable quantification of global DNA methylation – in just 2 hours

Product Description

The MethylFlash Global DNA Methylation (5‑mC) ELISA Easy Kit (Colorimetric) is a complete, ready‑to‑use solution for quantifying global 5‑methylcytosine (5‑mC) directly from purified DNA using a simplified one‑step ELISA‑like protocol. It combines high specificity for 5‑mC with a robust colorimetric readout, allowing you to determine global DNA methylation levels across a wide range of species and sample types. 

Based on EpigenTek’s fourth‑generation global DNA methylation technology, this Easy Kit further improves speed, simplicity, sensitivity, and reproducibility over previous MethylFlash formats and traditional methods such as LUMA, LINE‑1, Alu or LTR‑based assays. 

Key Features & Benefits

Fast – complete assay in about 2 hours
Reduced hands‑on steps mean you can go from DNA to quantitative 5‑mC data in roughly two hours, significantly shorter than many competing global methylation assays. 

Sensitive – detect very low methylation levels
The assay can detect as little as 0.05% methylated DNA using 100 ng of input DNA, enabling reliable measurement even in samples with low global methylation. 

Highly specific for 5‑mC
Capture and detection antibodies show high specificity for 5‑mC with no cross‑reactivity to unmethylated cytosine and no detectable cross‑reactivity to 5‑hydroxymethylcytosine within the specified sample range. 

Universal – any species, many sample types
The kit is designed for DNA from virtually any species (mammals, plants, fungi, bacteria, viruses) and accepts DNA derived from cultured cells, fresh or frozen tissues, paraffin‑embedded tissues, plasma/serum, and other body fluids. Both single‑stranded and double‑stranded DNA from ~200 bp up to full length are suitable. 

Robust & reproducible
An optimized buffer system and improved reagent formulation provide a larger signal window and reduced variation between replicates, supporting confident comparison of methylation levels across samples and experiments. 

Simple workflow – no DNA denaturation or plate blocking
The inherently low background of the assay eliminates the need for DNA denaturation and blocking steps, simplifying the protocol and minimizing opportunities for error. 

Absolute and relative quantification
Included positive and negative controls (synthetic methylated and unmethylated DNA) allow you to generate a standard curve (0.1–5%) and quantify both absolute amounts and relative differences in global methylation. 

Flexible format for low‑ to high‑throughput
The strip‑well 96‑well plate format supports both manual processing and automation, making it easy to scale from a few samples to larger screens. 

Optimized for Paired Use with 5-hmC

The kit is specifically optimized to be used alongside the MethylFlash Global DNA Hydroxymethylation (5‑hmC) ELISA Easy Kit (Colorimetric), enabling parallel quantitation of both methylated and hydroxymethylated DNA from your samples. 

How It Works

ELISA‑like 5‑mC detection directly from bound DNA

1. DNA is bound to specially treated strip‑wells with high DNA affinity.
2. Methylated cytosines (5‑mC) are recognized by a highly specific 5‑mC capture antibody followed by a detection system.
3. A colorimetric signal is developed and measured at 450 nm; the optical density is proportional to the percentage of methylated DNA in the sample.

Using the included positive and negative controls, a standard curve (0.1–5% 5‑mC) can be generated to calculate the absolute amount of methylated DNA and to compare methylation levels between different samples or experimental conditions. 

Sample & Input Requirements

Sample types

• Cultured cells (flask or microplate)
• Fresh or frozen tissues
• Paraffin‑embedded tissues
• Plasma/serum and other body fluids
• DNA isolated from any species including mammals, plants, fungi, bacteria, and viruses

Input DNA

• Recommended input: 100 ng DNA per well (optimized for best quantification)
• Acceptable range: 5–200 ng DNA per reaction
• DNA purity: A260/280 > 1.6, diluted in water or 1× TE (pH 7.5–8.0)
• DNA size: ~200 bp to full length; single‑ or double‑stranded DNA are suitable

Applications

Ideal for:

• Global DNA methylation profiling across tissues, cell lines, or treatment conditions
• Biomarker and disease‑related studies where hypomethylation or hypermethylation is of interest (e.g., oncology, toxicology, environmental epigenetics)
• Comparing global methylation between species (mammals, plants, fungi, bacteria, viruses)
• Screening for compounds or conditions that alter global 5‑mC levels
• Complementary analysis with 5‑hmC quantification using the paired 5‑hmC ELISA Easy Kit

Performance vs. Competing Kits

Independent method comparisons highlight several advantages of the MethylFlash 5‑mC ELISA Easy Kit over other global methylation kits:

A substantially higher number of published citations use EpigenTek’s technology.

Performance Data

Fig.1. Example of an optimal standard curve generated with 5-mC standard control.

Fig.2. Accurate quantification of 5-mC content of various DNA samples from different species using the MethylFlash Global DNA Methylation (5-mC) ELISA Easy Kit (Colorimetric). The results are closely correlated with those obtained by HPLC-MS.

Get started today

Contact us to get started with your quantification of global DNA methylation in just 2 hours.