Zymo Reagents

<h2>What are 5-hmC ELISA kits?</h2>
&nbsp;
5-hmC ELISA kits are antibody-based assays formulated specifically to quantify 5-hydroxymethylcytosine, the oxidized derivative of 5-methylcytosine produced by TET family enzymes, in a genomic DNA sample. The assay typically uses denatured single-stranded DNA coated on a plate well, recognized by an anti-5-hmC primary antibody and developed with an HRP-conjugated secondary antibody for a colorimetric readout proportional to the global level of 5-hmC in the sample.
&nbsp;
<h2 dir="ltr">What you will find:</h2>
&nbsp;
<ul>
<li dir="ltr" role="presentation">DNA hydroxymethylation ELISA assay kits for 5-hmC levels quantification</li>
<li dir="ltr" role="presentation">DNA methylation ELISA kits for comparative gene regulation analysis and measurement of 5-mC&nbsp;</li>
<li dir="ltr" role="presentation">DNA standards for accurate methylation and hydroxymethylation quantification&nbsp;</li>
<li dir="ltr" role="presentation">Control DNA sets and assay controls for experimental accuracy, validation, and reproducibility</li>
<li dir="ltr" role="presentation">HRP-based antibodies for the specific detection in ELISA workflows</li>
<li dir="ltr" role="presentation">ELISA coating and reaction buffers for enhanced assay performance and setup</li>
<li dir="ltr" role="presentation">Signal development reagents for quantitative readout and chemiluminescent or colorimetric detection</li>
</ul>
&nbsp;
<h2>How to use 5-hmC ELISA kits</h2>
&nbsp;
Confirm 5-hmC is detectable at relevant levels in your cell type or tissue
5-hmC is most abundant in neurons and embryonic stem cells, where it can constitute up to 20% of total modified cytosines in neuronal DNA, while most cultured cell lines have much lower 5-hmC levels that may fall below a plate-based antibody assay's reliable detection limit. Checking published 5-hmC abundance for your specific cell type helps confirm whether ELISA-level detection is appropriate before committing samples to the assay.
Use a set of mixed methylation standards to build a calibration curve
Running defined mixtures of 5-hmC-containing and unmethylated DNA at known ratios generates a standard curve that lets you convert the raw absorbance signal to an actual global hydroxymethylation percentage, since the assay signal alone cannot be interpreted as an absolute level without this calibration.
Confirm your antibody's specificity for 5-hmC versus 5-mC
Anti-5-hmC antibodies used in these kits are selected for specificity to 5-hydroxymethylcytosine rather than 5-methylcytosine, but published cross-reactivity data for the specific antibody in your kit is worth checking, particularly when studying sample types where both marks are present at similar levels.
Plan your experimental design to include appropriate negative and positive controls
Including a negative control (unmethylated DNA, expected to give background-level signal) and a positive control (5-hmC-containing DNA of known level) in every plate confirms the assay is performing as expected and catches plate-to-plate variability before it affects result interpretation.
Understand ELISA gives global level, not locus-specific 5-hmC information
A 5-hmC ELISA measures the average hydroxymethylation level across all DNA in the sample; it does not reveal which specific CpG sites carry 5-hmC, which requires hMeDIP-seq or TAB-seq for regional or base-level resolution respectively.
&nbsp;
<h2>Specifications context</h2>
&nbsp;
Anti-5-hmC antibodies used in ELISA formats are validated specifically for 5-hydroxymethylcytosine and selected for minimal cross-reactivity with 5-methylcytosine, since distinguishing these two modifications is the primary purpose of a dedicated 5-hmC assay rather than a general methylation assay. As of 2026, 5-hmC ELISA kits remain a standard first-pass tool in studies where many samples need to be screened for differences in global hydroxymethylation level before committing a smaller subset to high-resolution sequencing methods.
<a href="https://mbpinc.net/contact-us">Contact the expert team</a> at Molecular Biology Products Inc. (MBP) and find reliable ELISA kits and reagents for your lab.

ELISA Kits & Reagents

MSDS

Control DNA Set (5 x 40 µl)

D5425-5-C by Zymo Research is the Control DNA Set (5 × 40 µl), providing calibration standard DNA for use with the Quest 5-hmC DNA ELISA Kit to detect and quantify 5-hydroxymethylcytosine (5-hmC) in genomic DNA. Required for generating the ELISA calibration curve. For research use only.

Manufacturer

Shipping Condition

Storage Condition

Control E (100 ng/ul) (40 µl)

D5425-5-5 by Zymo Research is Control E, a mouse tissue-derived genomic DNA at 100&nbsp;ng/&micro;l (40&nbsp;&micro;l) with defined %5-hmC for standard curve construction in the Quest 5-hmC DNA ELISA Kit (D5425). Used alongside Control C (D5425-5-3). Research use only.

Plates

Conversion Plate w/ Cover Foil (2 Plates/Foils)

C2005 by Zymo Research is a Conversion Plate with Cover Foil for 96-well bisulfite conversion workflows. Each unit includes 2 non-skirted PCR plates (up to 150 µl per well) with 2 piercing-compatible cover foils. Compatible with most standard thermal cyclers.

<h2>What are epigenetics enzymes and inhibitors?</h2>
&nbsp;
Epigenetics enzymes and inhibitors are the molecular tools for directly studying or modulating the enzymatic machinery that writes, erases, or reads epigenetic marks. The major classes include: DNA methyltransferases (DNMT1, DNMT3A, DNMT3B), which catalyze the addition of a methyl group to cytosine; TET family enzymes (TET1, TET2, TET3), which oxidize 5-methylcytosine to 5-hydroxymethylcytosine; histone acetyltransferases (HATs) and histone deacetylases (HDACs), which add and remove acetyl groups from histone lysine residues; and histone methyltransferases (HMTs) including EZH2, DOT1L, and G9a. Small-molecule inhibitors of HDAC and DNMT enzymes are the most clinically and experimentally advanced of these tools.
&nbsp;
<h2 dir="ltr">What you will find:</h2>
&nbsp;
<ul>
<li dir="ltr" role="presentation">Micrococcal nuclease for nucleosome mapping and controlled chromatin digestion applications</li>
<li dir="ltr" role="presentation">DNA shearing enzymes for consistent fragmentation of DNA in sequencing and epigenetic workflows</li>
<li dir="ltr" role="presentation">DNA degradation enzymes for controlled breakdown and analysis of nucleic acids&nbsp;</li>
<li dir="ltr" role="presentation">CpG methylase enzymes for targeted DNA methylation (CpG-specific )&nbsp; in epigenetic modification studies</li>
<li dir="ltr" role="presentation">5-hmC glucosyltransferase for detection and labeling of hydroxymethylcytosine in DNA</li>
<li dir="ltr" role="presentation">SAM (S-adenosylmethionine) cofactors for supporting enzyme activity and methylation reactions</li>
<li dir="ltr" role="presentation">DNA digestion enzymes for genomic DNA processing workflows</li>
<li dir="ltr" role="presentation">dsDNA nuclease systems for controlled cleavage and analysis of double-stranded DNA</li>
</ul>
&nbsp;
<h2>How to choose epigenetics enzymes and inhibitors</h2>
&nbsp;
Choose DNMT inhibitors to globally reduce or deplete DNA methylation in cells
Nucleoside analog DNMT inhibitors like 5-azacytidine and decitabine incorporate into DNA and trap DNMT1, preventing maintenance methylation and causing passive demethylation through DNA replication, while newer non-nucleoside DNMT inhibitors are in development for more selective, non-genotoxic methylation reduction.
Choose HDAC inhibitors to globally increase histone acetylation
HDAC inhibitors including pan-HDAC hydroxamates like vorinostat (SAHA) or class-selective compounds reduce histone deacetylase activity and cause hyperacetylation of histone lysine residues, broadly promoting gene expression at silenced loci. HDAC inhibitors are the most clinically advanced class of epigenetic therapeutics, with multiple FDA-approved compounds.
Use TET enzyme preparations for in vitro 5-hmC oxidation studies
Purified TET1, TET2, or TET3 enzyme preparations allow controlled in vitro oxidation of 5-methylcytosine to 5-hydroxymethylcytosine and further oxidation products, useful for generating defined 5-hmC-containing substrate DNA or studying TET enzyme activity and selectivity.
Choose EZH2 or other HMT inhibitors for studying PRC2-dependent gene silencing
EZH2, the catalytic subunit of polycomb repressive complex 2 (PRC2), catalyzes H3K27 trimethylation (H3K27me3), a histone modification associated with repression of tumor suppressor genes, and EZH2 inhibitors like tazemetostat are used in research to probe the role of this silencing mark.
Combine inhibitors to study epistatic relationships between epigenetic marks
Because DNA methylation, histone acetylation, and histone methylation are mechanistically linked, combining HDAC inhibitors with DNMT inhibitors can produce synergistic effects on gene expression exceeding either alone, and these combination approaches are both experimental tools and the basis for combination epigenetic therapy research.
&nbsp;
<h2>Specifications context</h2>
&nbsp;
HDAC and DNMT inhibitors are the most clinically advanced classes of epigenetic therapeutics, with multiple HDAC inhibitors including vorinostat and romidepsin approved for hematological malignancies, EZH2 inhibitors including tazemetostat having received accelerated approval, and multiple BET inhibitors in clinical trials. As of 2026, CRISPR/dCas9 systems fused with DNMTs, TETs, and HDACs enable locus-specific epigenome editing, extending classic enzyme and inhibitor approaches toward targeted epigenetic modulation.
<a href="https://mbpinc.net/contact-us">Contact the expert team at MBP</a> and find the suitable enzymes and inhibitory reagents for your lab.

Enzymes & Inhibitors

Protocol

CpG Methylase (M. Sssl)

CpG Methylase (M. SssI) (E2010) by Zymo Research — recombinant CpG methyltransferase that completely methylates all 5′-CpG-3′ sites in dsDNA. Supplied with 10X buffer and 20X SAM. Available in 200 U and 400 U; optimal at 30°C; heat-inactivatable at 65°C.

Additional Information

Assay Condition

Heat Inactivation

Source

Storage

Unit Definition

CpG Methylase (M. Sssl) (200 U)

CpG Methylase (M. Sssl) (400 U)

Cytosine DNA Standard (2 µg)

D5405-1 by Zymo Research is the Cytosine DNA Standard (2 µg) containing 100% unmodified cytosines (no 5-mC, no 5-hmC), used to anchor the zero-modification point on calibration curves in 5-mC and 5-hmC ELISA quantitation assays. Compatible with Zymo Research 5-mC DNA ELISA Kit (D5325) and the 5-Methylcytosine &amp; 5-Hydroxymethylcytosine DNA Standard Set (D5405). Store at −20 °C. Made in USA.

<h2>What are DNA Cleanup Kits?</h2>
&nbsp;
DNA cleanup kits purify and concentrate pre-extracted or pre-amplified DNA by removing enzymes, salts, organic solvents (phenol, chloroform), unincorporated nucleotides, RNA, detergents, and other contaminants from DNA solutions using silica-membrane spin columns with a bind-wash-elute protocol. Unlike PCR cleanup kits that are specifically optimized to remove primers and dNTPs from PCR reactions, DNA cleanup kits accept broader input chemistries - including post-phenol-chloroform extraction DNA, genomic DNA in high-salt lysis buffers, restriction digest reactions, and dilute DNA solutions requiring concentration - and elute DNA in minimal volumes (as little as 6 ul) for immediate use in downstream applications. Use a DNA cleanup kit whenever your DNA preparation contains inhibitors incompatible with downstream enzymatic reactions, when residual phenol must be removed, or when dilute DNA must be concentrated into a small volume.
&nbsp;
<h2 dir="ltr">What you will find:</h2>
&nbsp;
<ul>
<li dir="ltr" aria-level="1">
DNA Clean &amp; Concentrator&trade; (DCC): Efficient spin-column systems (5, 25, 100, 500 preps) created to concentrate low-concentration DNA into ultra-pure, transfection-quality volumes.
</li>
<li dir="ltr" aria-level="1">
Genomic DNA Clean &amp; Concentrator: Tailored procedures for extracting large templates while maintaining genomic integrity for long-read sequencing uses.
</li>
<li dir="ltr" aria-level="1">
Select-a-Size DNA Clean &amp; Concentrator&trade;: Tailored systems for the recovery of size-specific fragments, efficiently eliminating adapter dimers and non-target amplicons.
</li>
<li dir="ltr" aria-level="1">
ZR-96 DNA Cleanup Systems: Scalable 96-well and MagBead configurations designed for high-throughput, automated processes while ensuring reproducibility-validated outcomes.
</li>
</ul>
&nbsp;
<h2>How to Choose a DNA Cleanup Kit</h2>
&nbsp;
Capacity and Input Volume
Zymo DNA Clean &amp; Concentrator kits are available in formats matched to DNA input mass: DCC-5 (&lt;=5 ug input, elute &gt;=6 ul), DCC-25 (&lt;=25 ug input, elute &gt;=25 ul), DCC-100 (&lt;=100 ug input, elute &gt;=25 ul). Choose the format whose maximum capacity exceeds your expected DNA input by at least 2-fold to avoid column overloading, which reduces yield and purity. For genomic DNA preparations from tissue (typically 5-30 ug), DCC-25 or DCC-100 is appropriate.
Minimum Elution Volume and Concentration
For maximum DNA concentration, Zymo DCC-5 elutes in 6 ul - the industry minimum for standard silica columns. If the input volume is 500 ul (1 ng/ul) and the elution is 6 ul, the final concentration reaches approximately 80 ng/ul (assuming 95% recovery), a concentration suitable for transfection, electroporation, and microinjection.
Size Selectivity
Standard DNA cleanup kits retain all DNA &gt;=100 bp. If size selection is also required, use gel extraction (for specific band isolation) or AMPure XP beads (for size-range selection by bead: volume ratio adjustment) rather than a standard cleanup kit.
Post-Phenol-Chloroform Cleanup
For DNA recovered from phenol-chloroform extraction, add 5 volumes of binding buffer and 2 volumes of 100% ethanol to the aqueous phase before column loading. The organic solvent in residual phenol or chloroform does not significantly impair column binding at these dilutions. Multiple column reloads allow processing of large aqueous-phase volumes through a single column.
RNase A Treatment for DNA-Only Preparations
For gDNA preparations that must be RNA-free for optical mapping or certain long-read sequencing library kits, add RNase A (100 ug/ml) to the DNA solution, incubate at 37 degrees C for 10 minutes, then proceed directly to DCC cleanup. The RNase digestion products are removed during the wash steps.
&nbsp;
<h2>Specifications Context</h2>
&nbsp;
Zymo DNA Clean &amp; Concentrator kits use a single-buffer system with Zymo-Spin column technology and recover DNA from PCR, enzymatic reactions, phenol-chloroform extractions, and dilute solutions. DNA is eluted in volumes as small as 6 ul, providing up to 80-fold concentration from 500 ul inputs. The kits remove enzymes, proteins, salts, organic solvents, dyes, unincorporated nucleotides, primers, and other small molecules, delivering DNA ready for restriction digestion, ligation, sequencing, Southern blotting, transfection, and long-read library preparation. DNA cleanup and concentration are standard workflow steps in genomics laboratories at Howard Hughes Medical Institute, Vanderbilt University, and MD Anderson Cancer Center, where removing trace inhibitors from gDNA before library preparation significantly improves NGS library complexity and uniformity.
&nbsp;
Enhance your capabilities&mdash;<a href="https://mbpinc.net/contact-us">reach out to the MBP Inc. team</a> now to obtain a quote for your DNA concentration requirements.

DNA Cleanup Kits

Brochure

DNA Clean & Concentrator Magbead Kit (100 preps)

D4012 by Zymo Research is a 100-prep magnetic bead DNA cleanup kit compatible with any open liquid handler or magnetic bead mover. Recovers DNA ≥25 bp (up to 150 kb HMW); &gt;90% recovery for ≥100 bp fragments; purity A260/A280 &amp; A260/A230 ≥1.8; elution in 30–50 µl. Suitable for PCR cleanup, NGS library prep, restriction digests, and automated high-throughput purification.

Binding Capacity

Purity

Sample Sources

Size

DNA Clean & Concentrator-100

The DNA Clean &amp; Concentrator-100 by Zymo Research purifies up to 100 µg of DNA from PCR, large restriction digests, or cell-free lysates in &lt;20 min, with 70–95% recovery for 50 bp–10 kb and ≥150 µL elution. Available as D4029 (25 Preps.) and D4030 (50 Preps.).

Detergent Tolerance

Elution Volume

Equipment

Sample Source

Size Range

Yield

DNA Clean & Concentrator™-100 (25 Preps.)

DNA Clean & Concentrator™-100 (50 Preps.)

DataSheet

DNA Clean & Concentrator-25

Zymo Research DNA Clean &amp; Concentrator-25: 25 µg binding, ≥25 µl elution, ~2 min protocol, 70–90% recovery (50 bp–10 kb). D4005 (50 preps) and D4006 (200 preps).

DNA Clean & Concentrator™-25 (50 Preps.)

DNA Clean & Concentrator™-25 (200 Preps.)

Zymo Research DNA Clean &amp; Concentrator-25: 25 µg binding capacity, ≥25 µl elution, ~2 min protocol, 70–90% recovery (50 bp–10 kb). Two sizes: D4033 (50 preps) and D4034 (200 preps).

DNA Clean & Concentrator-500

The DNA Clean &amp; Concentrator™-500 by Zymo Research purifies and concentrates up to 500 µg of DNA from large-scale restriction digests, crude preparations, and high-mass PCR reactions in under 20 minutes, with 70–95% recovery for 50 bp–10 kb fragments. Available as D4031 (10 Preps.) and D4032 (20 Preps.).

DNA Clean & Concentrator™-500 (10 Preps.)

DNA Clean & Concentrator™-500 (20 Preps.)

DNA Clean & Concentrator™-5

DNA Clean & Concentrator-5 by Zymo Research — 2-minute cleanup; ≥6 µL elution; 50 bp–23 kb; ≤5 µg binding capacity; A260/A280 > 1.8; removes enzymes, dNTPs, and buffers from PCR and enzymatic reactions. Available in 10/50/200-prep kits (uncapped or capped columns), or individual Binding and Wash buffers.

DNA Clean & Concentrator™-5 (50 Preps.) (Uncapped)

DNA Clean & Concentrator™-5 Kit (10 Preps.)(Uncapped)

DNA Clean & Concentrator™-5 (200 Preps.)(Uncapped)

DNA Clean & Concentrator™-5 (50 Preps.) (Capped)

DNA Clean & Concentrator™-5 (200 Preps.) (Capped)

DNA Binding Buffer (25 ml)

DNA Binding Buffer (50 ml)

DNA Wash Buffer (Concentrate) (24 ml)

DNA Wash Buffer (Concentrate) (48 ml)

DNA Wash Buffer (Concentrate) (6 ml)

DNA Binding Buffer (100 ml)

<h2>What are ELISA kits for methylation in this category?</h2>
&nbsp;
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.
&nbsp;
<h2 dir="ltr">What you will find:</h2>
&nbsp;
<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>
&nbsp;
<h2>How to choose methylation ELISA kits and reagents</h2>
&nbsp;
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.
&nbsp;
<h2>Specifications context</h2>
&nbsp;
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.

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.