ELISA Kits & Reagents

<h2>What are ELISA kits for methylation in this category?</h2>
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ELISA kits for DNA and RNA methylation in this category are antibody-based assays that quantify the global level of a specific epigenetic mark, most commonly 5-methylcytosine in genomic DNA, 5-hydroxymethylcytosine in genomic DNA, or N6-methyladenosine in RNA, using a plate-based immunoassay format without requiring bisulfite conversion or sequencing. Denatured, single-stranded DNA or RNA from a sample is coated on a plate well, and a mark-specific primary antibody followed by an HRP-conjugated secondary antibody generates a colorimetric signal proportional to the level of the target modification.
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<h2 dir="ltr">What you will find:</h2>
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<ul>
<li dir="ltr" role="presentation">5-mC DNA ELISA kits for quantitative analysis of DNA methylation&nbsp;</li>
<li dir="ltr" role="presentation">DNA controls for validation and calibration of the assay to ensure experimental&nbsp; reproducibility and accuracy&nbsp;</li>
<li dir="ltr" role="presentation">Secondary antibody reagents for specific signal generation and detection in ELISA assays</li>
<li dir="ltr" role="presentation">Coating buffers for consistent assay performance and optimized plate preparation&nbsp;</li>
</ul>
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<h2>How to choose methylation ELISA kits and reagents</h2>
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Match your kit to the specific epigenetic mark you're quantifying
Different kits use antibodies against specific modifications: 5-methylcytosine ELISA kits quantify global DNA methylation, 5-hmC ELISA kits quantify global hydroxymethylation, and m6A ELISA kits quantify RNA N6-methyladenosine. Using a kit with the wrong antibody specificity will not report the mark you intend to measure.
Use methylation standards to build a calibration curve for quantitative results
Running a series of methylation standards with defined percentages alongside your samples allows the absorbance readout to be converted to a specific, meaningful methylation percentage, since the raw ELISA signal alone cannot be interpreted as absolute methylation level without a calibration.
Understand that ELISA measures global levels, not locus-specific methylation
A methylation ELISA gives the overall average percentage of methylation across the genome in a sample, which is useful for comparing broad methylation changes between conditions but cannot identify which specific genes or regions are differentially methylated the way bisulfite sequencing can.
Confirm the required DNA denaturation step is appropriate for your sample type
ELISA kits for DNA methylation typically require single-stranded, denatured DNA coated directly on the plate, and confirming this denaturation step is properly performed is necessary for consistent antibody binding and reproducible signal across wells.
Be aware of cross-reactivity with similar modifications when interpreting results
An antibody against 5-methylcytosine may have some cross-reactivity with 5-hydroxymethylcytosine, and confirming the antibody's published cross-reactivity data helps interpret whether your signal specifically reflects 5-mC, 5-hmC, or both, which matters for studies in cell types where 5-hmC is abundant.
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<h2>Specifications context</h2>
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The 5-methylcytosine ELISA workflow uses an indirect ELISA methodology where denatured, single-stranded DNA samples are coated directly on the plate well surfaces and a 5-mC monoclonal antibody and an HRP-conjugated secondary antibody are added sequentially, generating a colorimetric signal proportional to the total level of methylated cytosine in the sample. As of 2026, methylation ELISA kits are particularly valued for high-throughput screening studies comparing global methylation levels across many treatment conditions or patient groups where single-base-resolution data is not the primary goal.
This category sits within DNA/RNA Methylation, alongside Methylation Standards for calibration, and is related to DNA Hydroxymethylation and Non-Bisulfite Kits &amp; Reagents for higher-resolution methylation analysis. See the full reagents catalog and use Quick Order for recurring orders.
<a href="https://mbpinc.net/contact-us">Contact the expert team of&nbsp; Molecular Biology Products Inc. (MBP)</a> to get high-quality ELISA Kits and Reagents for your lab.

DataSheet

D5325-1-15/D5325-1-30 - MSDS

MSDS

5-mC Coating Buffer

5-mC Coating Buffer and 5-mC ELISA Buffer by Zymo Research — components of the 5-mC DNA ELISA Kit (D5325, D5326). Coating Buffer available in 15mL (D5325-1-15) or 30mL (D5325-1-30); ELISA Buffer in 250mL (D5325-2-250). Stored at 4°C.

Manufacturer

Shipping Condition

Storage Condition

5-mC Coating Buffer (15 ml)

5-mC Coating Buffer (30 ml)

5-mC ELISA Buffer (250 ml)

Protocol

5-mC DNA ELISA Kit (2 x 96 Preps.)

D5326 by Zymo Research is a 5-mC DNA ELISA Kit for colorimetric quantification of global 5-methylcytosine in purified DNA. Accepts 10–200 ng DNA per well, detects methylation above 0.5% 5-mC, and completes in under 3 hours. Kit contains reagents for 2 × 96 reactions.

Detection

Equipment

Input

Sample Source

5-mC DNA ELISA Kit (96 Preps.)

The 5-mC DNA ELISA Kit (D5325) by Zymo Research quantifies global 5-methylcytosine content in 96 samples using a colorimetric anti-5-mC antibody assay (Clone 7D21) with ≥0.5% detection sensitivity, a 7-point standard curve, and a total assay time under 3 hours.

D5325-5-1-MSDS

D5325-5-2-MSDS

DNA Controls

DNA Controls are available in the following variants: D5325-5-1, D5325-5-2.DNA Controls D5325-5-1 by Zymo Research — Negative Control (0% 5-mC) for the 5-mC DNA ELISA Kit. E. coli genomic dsDNA at 100 ng/µl, 50 µl per unit, stored at −20°C. Used for standard curve construction and assay baseline in global DNA methylation quantification.

Negative Control (100 ng/µl) (50 µl)

Positive Control (100 ng/µl) (50 µl)

Secondary Antibody

Zymo Research HRP-conjugated Secondary Antibody (1 µg/µl) for 5-mC DNA ELISA Kit (D5325/D5326): detects anti-5-mC Clone 7D21 at 405 nm. Two volumes: D5325-3-15 (15 µl) and D5325-3-30 (30 µl).