Nucleosome Elisa

Nucleosome ELISA is an enzyme-linked immunosorbent assay that detects and quantifies nucleosomes, complexes of DNA wrapped around histone proteins, in biological samples. The method uses antibodies to capture nucleosomes on a coated microplate, while a second enzyme-linked antibody binds the captured complexes; adding a substrate produces a measurable color signal proportional to nucleosome abundance. This assay provides a sensitive, scalable approach for studying chromatin fragmentation and cell death, including apoptosis, and can support research into tissue injury, disease mechanisms, and therapeutic responses. Its quantitative readout also enables comparisons across experimental conditions and biological samples.

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Research

JoVE Journal - Biology
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Detection of Post-translational Modifications on Native Intact Nucleosomes by ELISA

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Cited by 8 •

2011

Nucleosome ELISA (NU-ELISA) is a sensitive and quantitative method to detect global patterns of post-translational modifications in preparations of native, intact nucleosomes. These modifications include methylations, acetylations, and phosphorylations at specific histone amino acid residues, and hence NU-ELISA provides a global proteomic assay of the overall chromatin modification states of specific cell types.

Education

JoVE Core - Molecular Biology

The Nucleosome

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2020

DNA in a human cell is almost 2m long and it is packed inside a tiny nucleus that is only a few microns in diameter. The level of compaction of DNA inside the nucleus is astonishing. It is organized into several sequentially higher levels of compaction to fit into such a tiny space. The most compact form of DNA is a chromosome that can be seen under a microscope in a dividing cell. DNA is wound twice around a protein complex called histone core, that consist of 8 histone proteins. This complex...

Nucleosome Remodeling

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2020

Nucleosomes are the basic units of chromatin compaction. Each nucleosome consists of the DNA bound tightly around a histone core, which makes the DNA inaccessible to DNA binding proteins such as DNA polymerase and RNA polymerase. Hence, the fundamental problem is to ensure access to DNA when appropriate, despite the compact and protective chromatin structure. Nucleosome remodeling complex Eukaryotic cells have specialized enzymes called ATP-dependent nucleosome remodeling enzymes. These enzymes...

The ELISA Method

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2023

An enzyme-linked immunosorbent assay (ELISA) is typically performed to detect the presence and/or amount of a target protein of interest within an experimental sample. Detection of the target protein is made possible by antibodies, which make the ELISA an immunoassay. Through a series of incubation and washing steps, these antibodies, which are frequently linked, or conjugated, to an enzyme, will detect protein coating the bottom of a well on a microtiter plate. When exposed to a substrate,...

The Nucleosome Core Particle

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2020

Nucleosomes are the DNA-histone complex, where the DNA strand is wound around the histone core. The histone core is an octamer containing two copies of H2A, H2B, H3, and H4 histone proteins. The paradox Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their main responsibility is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. While on the other hand, they must allow polymerase enzymes to access DNA...

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