EpiStart DNA Methylation (5-mC) ELISA Kit

The EpiStart™ DNA Methylation (5-mC) ELISA Kit is a colorimetric ELISA-based assay designed to detect and quantify global DNA methylation. The kit measures 5-methylcytosine (5-mC) levels in genomic DNA from virtually any species and enables calculation of unknown methylation levels in your DNA samples using a standard curve.

Detailed Description:
This kit provides a streamlined, hands-on ELISA workflow for analyzing DNA methylation. The assay is performed in three simple steps:

• DNA Binding – DNA samples and controls bind to high-affinity assay wells
• Methylated DNA Detection – A 5-mC-specific antibody-based system detects methylated cytosine
• Color Development & Measurement – Color intensity is measured and used to quantify %5-mC

Students can quantify global DNA methylation using a microplate reader or compatible analysis tools, reinforcing both experimental technique and data interpretation.

Background
Epigenetic modifications are like “on/off switches,” controlling when and where genes are expressed. The most widely studied and best characterized is 5-methylcytosine (5-mC) in DNA. 5-mC occurs when a methyl group is covalently added at the 5-carbon position of cytosine nucleotides. This has the effect of inhibiting transcription, the process of copying DNA into RNA. DNA methylation is a natural and reversible mechanism that instructs a gene at what times and at which locations in the body it should be turned either on or off. When the DNA methylation machinery stops functioning properly, this can lead to the development of numerous diseases, including cancer. Methylated DNA has also proven quite useful in various clinical applications as a biomarker for particular medical conditions.

Principle & Procedure
This kit contains all reagents necessary for the quantification of global DNA methylation. In this assay, DNA is bound to strip-wells with high DNA affinity. The methylated fraction of DNA (5-mC) is detected using a 5-mC antibody-based detection complex and quantified colorimetrically by measuring absorbance with a microplate spectrophotometer. A standard curve is generated using known 5-mC controls, allowing accurate determination of methylation levels in unknown DNA samples from a wide range of species.

Key Benefits:

• Quantitative analysis: Calculate %5-mC using a standard curve
• Visual detection: Colorimetric readout correlates with methylation levels
• Complete kit: Includes controls, samples, and all required reagents
• Fast workflow: Completed in approximately 80–90 minutes
• Educational focus: Introduces ELISA and epigenetics concepts through hands-on learning
• Broad compatibility: Suitable for DNA from virtually any species

Educational Value:

• Understand DNA methylation and its role in gene regulation
• Perform and interpret a standard curve
• Apply ELISA-based techniques used in real research labs
• Analyze and interpret quantitative biological data

Aligned with AP Biology “Big Idea” IST 2.A.2.

Time Requirement:
Approximately 80–90 minutes, suitable for completion within one class + lab period.

Science & Engineering Practices:

• Planning and carrying out investigations
• Developing and using models
• Analyzing and interpreting data
• Obtaining, evaluating, and communicating information

Learning Objectives:

• Formulate and justify hypotheses about DNA methylation levels
• Quantify methylation using a colorimetric assay
• Interpret results using a standard curve
• Understand how epigenetic modifications influence gene expression
• Gain hands-on experience with ELISA-based techniques

Recommended Prerequisite Skills:

• Basic pipetting techniques

No FAQs found at this time