Zymo Reagents

<h2>What are ChIP buffers?</h2>
&nbsp;
ChIP buffers are the aqueous formulations used at each stage of a chromatin immunoprecipitation protocol: cell lysis buffer disrupts the plasma membrane and releases nuclear content, nuclear lysis buffer solubilizes chromatin under conditions compatible with subsequent immunoprecipitation, sonication buffer maintains chromatin stability during shearing, ChIP dilution buffer reduces SDS concentration from the lysis buffer to allow antibody-antigen binding, sequential wash buffers with increasing ionic strength and detergent remove non-specifically bound chromatin while preserving the specific antibody-antigen-DNA complex, and elution buffer releases the captured complex from the antibody-bead resin before de-crosslinking.
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<h2 dir="ltr">What you will find:</h2>
&nbsp;
<ul>
<li dir="ltr" role="presentation">MN Digestion Buffer used in MNase-based workflows for enzymatic fragmentation of&nbsp; chromatin&nbsp;</li>
<li dir="ltr" role="presentation">Chromatin Shearing Buffer using sonification for chromatin disruption and preparation</li>
<li dir="ltr" role="presentation">5X MN Stop Buffer for the termination of enzymatic digestion reactions</li>
<li dir="ltr" role="presentation">Chromatin Dilution Buffer used in ChIP assays to provide optimized conditions for antibody binding&nbsp;</li>
<li dir="ltr" role="presentation">Chromatin Wash Buffer I, II, and III for background reduction and step-by-step non-specific binding removal&nbsp;</li>
<li dir="ltr" role="presentation">ChIP DNA Binding Buffer for purification and capture of immunoprecipitated DNA</li>
<li dir="ltr" role="presentation">5X Chromatin Elution Buffer for protein&ndash;DNA complexes recovery</li>
<li dir="ltr" role="presentation">5M NaCl for enhancing elution and chromatin stabilization</li>
<li dir="ltr" role="presentation">Nuclei Prep Buffer used before chromatin extraction for superior nuclear isolation</li>
</ul>
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<h2>How to use ChIP buffers</h2>
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Always add protease inhibitors to lysis and wash buffers immediately before use
Protease inhibitors must be added fresh to cell lysis and nuclear lysis buffers immediately before use, since most inhibitors are unstable in aqueous solution and lose activity quickly, and without them, nuclear proteases released during lysis will degrade histone tails and reduce ChIP enrichment of histone modifications.
Confirm SDS concentration is diluted before immunoprecipitation
High SDS in the nuclear lysis buffer is necessary for chromatin solubilization and sonication, but must be diluted approximately 10-fold using ChIP dilution buffer before antibody addition, since high SDS disrupts antibody-antigen binding and reduces immunoprecipitation efficiency.
Keep all buffers and chromatin on ice throughout the workflow after lysis
Chromatin prepared for ChIP is sensitive to non-specific interactions and protease activity after cell lysis, and maintaining samples at 4&deg;C throughout all lysis, sonication, immunoprecipitation, and wash steps minimizes these effects and preserves specificity of the enrichment.
Optimize wash buffer stringency to balance specificity against yield
ChIP wash buffers are applied sequentially at increasing ionic strength and detergent concentration to remove non-specifically bound chromatin; conditions too stringent can also strip the specific antibody-antigen-DNA complex from the beads, reducing yield, so optimization of wash conditions for each antibody-target combination improves performance.
Confirm elution buffer composition fits your downstream application
Standard ChIP elution buffers containing SDS and sodium bicarbonate release the protein-DNA complex and reverse crosslinks, but high SDS in the eluate can inhibit some NGS library preparation enzymes, making a DNA cleanup step after elution important before proceeding to library prep.
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<h2>Specifications context</h2>
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Protease inhibitor cocktails, added fresh to lysis buffers immediately before ChIP, are a standard component across kit formats and in-house protocols, since even brief protease activity can compromise histone modification detection by degrading the histone tails that ChIP antibodies recognize. As of 2026, ChIP buffer formulations compatible with low-input chromatin and NGS library preparation continue to be optimized for primary cell and tissue ChIP workflows.
<a href="https://mbpinc.net/contact-us">Contact the expert team </a>at MBP and request a quote today to get high-quality ChIP buffers for your lab.

Buffers

MSDS

Chromatin Dilution Buffer (30 ml)

D5210-2-30 Chromatin Dilution Buffer by Zymo Research — 30 ml, room-temperature storage — dilutes ChIP-grade antibody during chromatin immunoprecipitation. Compatible with Zymo-Spin ChIP Kit (D5209, D5210).

Manufacturer

Shipping Condition

Storage Condition

Chromatin Shearing Buffer (30 ml)

D5210-1-30 by Zymo Research is a 30 ml chromatin shearing buffer for nuclei preparation and sonication in ChIP workflows, supporting reproducible fragment sizing to 100–600 bp. Sold as a standalone refill for the Zymo-Spin ChIP Kit (D5210).

Chromatin Wash Buffer I (30 ml)

D5210-3-30 by Zymo Research is Chromatin Wash Buffer I, 30 mL. Used as the first wash of Protein A bead/antibody/chromatin complexes in ChIP workflows. Compatible with the Zymo-Spin ChIP Kit (D5209, D5210). Sold separately. Store at room temperature.

Chromatin Wash Buffer II (30 ml)

D5210-4-30 by Zymo Research is Chromatin Wash Buffer II, a 30 ml buffer for the second chromatin wash step in the Zymo-Spin ChIP Kit (D5209, D5210) workflow. Stored at room temperature; available individually for kit buffer replenishment.

Chromatin Wash Buffer III (30 ml)

Chromatin Wash Buffer III (D5210-5-30) by Zymo Research is a 30 ml wash reagent for the third sequential wash step in Zymo-Spin ChIP Kit (D5209, D5210) protocols, removing residual non-specific binding from Protein A bead/antibody/chromatin complexes before DNA elution. Sold separately for kit replenishment or standalone use; store at room temperature. MBP is a registered vendor for Howard Hughes Medical Institute, Vanderbilt University, MD Anderson Cancer Center, Eurofins, and many other institutes across the USA.

<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

Coating Buffer

D5425-1-15 by Zymo Research — Coating Buffer, 15 mL; component of the Quest 5-hmC DNA ELISA Kit; used in the microplate coating step for 5-hydroxymethylcytosine (5-hmC) detection and quantification. For research use only.

Coating Buffer (15 ml)

Coating Buffer (30 ml)

Plates

Collection Plate (2 Plates)

C2002 by Zymo Research — 96-well round-bottom collection plate; 1.2 mL per well; clear polypropylene; compatible with Silicon-A, Zymo-Spin I-96, IB-96, and III-96 plates; sold as 2 plates per pack.

Spin Columns

Collection/Filter Assemblies

Collection/Filter Assemblies by Zymo Research support large-volume lysate processing in DNA and RNA purification workflows. Available as Zymo-Spin III-P (15 mL + 50 mL reservoirs, 5-pack, C1040-5) or Zymo-Spin III-S with LuerLock caps (15 mL reservoir, 25-pack, C1049-25).

Binding Capacity

Compatibility

Elution Volume

Format

Matrix/Construction

Volumetric

Zymo-Spin™ III-P Column Assembly w/ 15 ml and 50 ml Reservoir (5 Pack)

Zymo-Spin™ III-S Columns w/ LuerLock Caps and 15 ml Reservoir (25 Pack)

Control A (100 ng/ul) (40 µl)

D5425-5-1 by Zymo Research is Control A, a 40 µl aliquot of 100 ng/µl control DNA for use with the Quest 5-hmC DNA ELISA Kit for 5-hydroxymethylcytosine detection and quantification. Store at -20°C. Research use only.

Control B (100 ng/ul) (40 µl)

D5425-5-2 by Zymo Research is Control B, a calibrated mouse genomic DNA standard at 100 ng/µl (40 µl volume) for use as a reference point in the Quest 5-hmC DNA ELISA Kit (D5425/D5426) standard curve. Used to quantify 5-hydroxymethylcytosine (5-hmC) in unknown samples at ≥0.02% sensitivity per 100 ng DNA. Must be run on the same ELISA plate as unknowns. Store at −20 °C.

Control C (100 ng/ul) (40 µl)

D5425-5-3 by Zymo Research is Control C, 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). Research use only.

Control D (100 ng/ul) (40 µl)

D5425-5-4 Control D by Zymo Research — 100 ng/µl, 40 µl, 0.23% 5-hmC — intermediate calibration standard for the Quest 5-hmC DNA ELISA Kit standard curve. Store at -20°C.