Mcherry-tagged Histone H2b
Mcherry-tagged Histone H2b - Related Videos
02:32
Histone Modification
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2020
The histone proteins have a flexible N-terminal tail extending out from the nucleosome. These histone tails are often subjected to post-translational modifications such as acetylation, methylation, phosphorylation, and ubiquitination. Particular combinations of these modifications form “histone codes” that influence the chromatin folding and tissue-specific gene expression. Acetylation The enzyme histone acetyltransferase adds acetyl group to the histones. Another enzyme, histone deacetylase,...
02:30
Histone Variants at the Centromere
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2020
Histone variants are the histone proteins with structural and sequence variations. These variants may be regarded as “mutant” forms that replace their canonical histone counterparts in the nucleosomes. Specific post-translational modifications on the histone variants enable further chromatin complexity and regulate tissue-specific gene expression. The most common histone variants are from histone H2A, H2B, and linker histone H1 families. However, several variants of histone H3 variants are also...
09:26
Reconstitution of Nucleosomes with Differentially Isotope-labeled Sister Histones
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Cited by 1 •
2017
This protocol describes the reconstitution of nucleosomes containing differentially isotope-labeled sister histones. At the same time, asymmetrically post-translationally modified nucleosomes can be generated after using a premodified histone copy. These preparations can be readily used to study modification crosstalk mechanisms, simultaneously on both sister histones, by using high-resolution NMR spectroscopy.
10:54
Extraction of Histones from Clinical Specimens for Epigenetic Profiling by Mass Spectrometry
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2025
This protocol enables efficient histone extraction from various clinical samples for subsequent post-translational modification analysis by LC-MS/MS. By facilitating robust detection of key histone modifications, it serves as a valuable tool for comprehensive epigenetic profiling in large clinical cohorts.
09:47
Analysis of Histone Antibody Specificity with Peptide Microarrays
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Cited by 22 •
2017
This manuscript describes methods for applying peptide microarray technology to specificity profiling of antibodies that recognize histones and their post-translational modifications.
FAQs
The H2B portion associates with nucleosomal DNA, providing the targeting component that keeps the fusion protein with chromatin. This positioning allows the attached mCherry signal to follow nuclear material as its organization changes. Consequently, researchers can examine chromatin distribution and chromosome behavior through the fluorescent pattern rather than relying on repeated DNA staining.
The mCherry portion supplies detectable red fluorescence when the sample receives appropriate illumination. It does not provide the chromatin-targeting function; that role comes from H2B. Separating these roles explains why the fusion can convert chromatin-associated behavior into an optical readout, allowing nuclear structures and their changes to be followed in biological samples.
The fluorescent fusion can provide an ongoing chromatin-associated signal during observations, whereas DNA staining is not required at every observation. This distinction is especially relevant for live-cell imaging, where researchers may want to examine nuclear organization, chromosome condensation, or cell divisions over time without treating the sample anew for each viewing.
Cells first express the fusion protein, after which the biological sample is observed under illumination suitable for detecting mCherry. In live-cell settings, repeated observations can reveal nuclear organization, chromosome condensation, mitosis, and cell-cycle progression. In fixed samples, the same label supports visualization of chromatin and nuclei at the prepared observation point.
It is useful when an experiment requires nuclear or chromatin behavior to be followed over time, including cell divisions or lineage patterns. The approach can support comparisons across successive stages in the same biological system. It also remains relevant for fixed-sample imaging when the experimental design focuses on visualizing chromatin or nuclei at a defined point.
This label provides a readout for changes in nuclear organization, chromosome condensation, mitosis, and cell-cycle progression. During cell division, the chromatin-associated signal can help reveal how nuclear material is arranged across successive stages. Tracking divisions or lineage patterns extends the approach from individual nuclear images to observations of biological history over time.