MethylFlash Global DNA Methylation (5-mC) ELISA Easy Test (Colorimetric), 48 reactions

Biozoomer advice

• The measurement of the overall percentage or relative amount of 5-methylcytosine (5-mC) in genomic DNA
• Gentaur Epigenetics protocol
• Typical use cases:
  • Epigenetics research
  • Cancer biology
  • Developmental biology
  • Toxicology / environmental exposure studies
  • Drug or treatment-induced methylation changes
• Sample types: Purified genomic DNA from cells, tissues, blood, etc.

Assay Principle (ELISA-based, colorimetric)

1. DNA binding: Genomic DNA is immobilized onto strip-well plates.
2. Detection: A 5-mC-specific antibody binds methylated cytosines.
3. Signal development: A secondary antibody and colorimetric substrate generate a signal.
4. Quantification: Absorbance is measured by a microplate reader (typically at 450 nm).
5. Output: Relative or absolute global DNA methylation level, calculated using provided controls/standards.

Key Features

• Format: ELISA, colorimetric
• Throughput: 48 reactions
• Sensitivity: Detects as little as ~0.5% global methylation
• DNA input: Typically 50–100 ng per reaction
• Time to results: ~2–3 hours
• Equipment required: Standard ELISA plate reader (no fluorescence needed)

Advantages

• No bisulfite conversion required
• Faster and simpler than LC-MS or sequencing-based methods
• Suitable for comparative screening across many samples
• Good reproducibility for global methylation profiling

Limitations

• Measures global methylation only (not locus-specific)
• Does not distinguish between different genomic regions
• Semi-quantitative unless carefully calibrated with standards

Typical Output

• Absorbance values converted to:
  • % 5-mC of total DNA, or
  • Relative methylation levels compared to controls

When to Use This Kit

Use this assay if you need:

• Rapid screening of methylation changes
• Comparison between treatments or conditions
• A cost-effective alternative to sequencing-based methylation analysis

Do not use it if you need:

• CpG-site–specific resolution
• Genome-wide methylation maps

Publications KULeuven  with Gentaur MethylFlash:

MSDS