A designer nucleosome is reconstituted in vitro from recombinant histone octamers and defined DNA sequences. Specific PTMs are introduced in one of two ways. Chemically modified histones can be synthesised by expressed protein ligation. Alternatively, amber codon suppression can install non-natural amino acids. The result is a nucleosome population that is homogeneous for a single, precisely defined modification state. This differs from chromatin taken from cells, where multiple modification states coexist.
Epigenetics Research Tools
Recombinant Nucleosomes: dNucs, rNucs and Chromatin Remodelling Substrates
Recombinant nucleosomes with chemically defined histone modifications for precise chromatin assays
These recombinant nucleosomes give you an exact modification state to work with, removing the ambiguity of bulk chromatin prepared from cells. This confidence supports characterising chromatin-modifying enzymes or screening epigenetic drug candidates. The portfolio includes recombinant human mono- and dinucleosomes assembled on defined DNA templates, including 147 bp Widom 601-based formats. Depending on the product, substrates contain defined histone PTMs, mutations, variants, linker DNA or fluorescent labels.
The portfolio provides a broad range of defined histone PTMs, cancer-associated mutations, variants, methylated-DNA nucleosomes, chromatin-remodelling substrates and spike-in controls. Biotinylated and non-biotinylated versions are available, so you can run capture-based assays or free-in-solution assays.
Recombinant nucleosome monomers support your enzyme processivity studies. Dinucleosomes support linker-length studies. Tailless versions lack histone tail domains, so you can use them as controls for tail-independent binding studies. Chromatin remodelling substrates include FRET-based configurations that report nucleosome sliding in real time. These enable ATPase-coupled remodelling assays with SMARCA family enzymes.
Recombinant nucleosome spike-in panels use these designer nucleosomes for biochemical assays as carrier nucleosome controls. This links substrate biochemistry directly to your ChIP-seq or CUT&RUN quantification.
Products are supplied as purified, pre-assembled nucleosome substrates. They are ready for direct use in your pull-down or FRET workflows and as nucleosome substrates in your drug discovery assays, a step up from peptides that opens access to historically challenging targets.
Choosing the right recombinant nucleosome format
- If you need a substrate for a single histone modification state in a pull-down or enzyme assay: a modified designer nucleosome monomer in the modification state you need suits this format. It is available in both biotinylated and non-biotinylated formats, across more than 76 modification states.
- If you need minimal, tailless nucleosomes for structural studies or controls: tailless recombinant nucleosomes with linker DNA, where the histone tail domains have been removed, suit this format. These are useful for tail-independent binding studies.
- If you need dinucleosomes to study enzyme processivity or linker effects: biotinylated dinucleosomes with two nucleosomes separated by a defined linker DNA length suit this format. These support capture-based processivity assays.
- If you are measuring chromatin remodelling activity by FRET in real time: EpiDyne-FRET substrates provide a fluorescent readout of nucleosome-remodelling activity and are compatible with active remodelling complexes such as SMARCA4, SMARCA2 and ACF.
- If you need nucleosome spike-in controls for ChIP-seq or CUT&RUN normalisation: a recombinant nucleosome spike-in panel carrying defined modification states on key H3 residues suits this format. This allows quantitative normalisation of immunoprecipitation efficiency across your samples.
Parent category: epigenetics research kits
Applications
Chromatin enzyme kinetics
Defined modification-state nucleosomes serve as substrates for your histone-modifying enzyme assays, covering HATs, HDACs, HMTs and reader proteins.
Drug screening
Modified or unmodified nucleosomes presented to candidate bromodomain, chromodomain or Tudor domain inhibitors support your pull-down or competition assays with a known PTM context.
ChIP and CUT&RUN normalisation
Spike-in panels carrying defined H3K4 or H3K27 methylation levels, added to your ChIP and CUT&RUN reactions, support quantitative comparison of pulldown efficiency.
Chromatin remodelling studies
FRET-labelled nucleosome substrates measure real-time sliding, unwrapping or eviction by SWI/SNF, ISWI or CHD family remodellers in your system.
Structural biology
Homogeneous, modification-specific nucleosomes suit your cryo-EM or X-ray studies of chromatin complexes, avoiding the heterogeneity of bulk cellular chromatin.
Antibody specificity validation
A panel of modified nucleosomes tests whether your anti-PTM antibodies recognise their intended target, not adjacent or cross-reactive states.
Frequently asked questions
Tailless nucleosomes lack the flexible N-terminal histone tail domains. These tails are the primary substrates for PTM writers and readers. Researchers use tailless nucleosomes as negative controls, to confirm that an observed interaction or enzymatic activity depends on the tail and not the nucleosome core. They are also used in structural studies, where disordered tails would reduce resolution. This helps isolate core particle-mediated effects from tail-mediated effects.
When added at a known amount before immunoprecipitation, recombinant nucleosomes carrying a specific modification serve as an internal standard. The fraction of spike-in recovered in the pulldown reflects the efficiency of that experiment. Scaling sequencing read counts to spike-in recovery corrects for tube-to-tube variation in IP efficiency. Without this correction, you cannot quantitatively compare histone mark levels across conditions, time points, or cell types.
Yes. A panel of modified and unmodified recombinant nucleosomes gives the most stringent way to test whether an anti-PTM antibody recognises its intended target, and not adjacent or cross-reactive states. For example, you can test an anti-H3K27me3 antibody against nucleosomes carrying H3K27me1, H3K27me2, H3K27ac, and unmodified H3K27 in parallel. This defines selectivity before you commit to large-scale ChIP or immunofluorescence experiments.
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