Volition Nu.Q^® Discover H3.1 Research Use Only Kit

Nu.Q^® Discover H3.1 ELISA Research Use Only Kit

Our Nu.Q^® Discover H3.1 ELISA Research Use Only kit is a comprehensive array of specialized buffers and reagents meticulously formulated to detect quantitative levels of circulating intact nucleosomes containing Histone H3 in plasma.

The nucleosome core particle is composed of an octamer of histone proteins comprising a tetramer of histone H3 and H4 and two dimers H2A-H2B, around which 147bp of DNA is wrapped. In addition to the post-translational modifications occurring on the N-terminal tails of the histone, histone variants (for histone H3, H2A and H2B but not H4) could be incorporated. The canonical H3.1 histone variant is incorporated during S-phase, is maintained at high levels in cells dividing at a high rate but is massively evicted in cells undergoing their last cell cycle before exit to differentiation. During cellular damage, such as apoptosis or necrosis, chromatin is fragmented into oligo- or mono- nucleosomes which can be released into the blood stream. Histones and DNA are subject to a variety of epigenetic modifications.

The Nu.Q^® Discover H3.1 ELISA Research Use Only kit is a powerful research tool for a wide range of research areas that study nucleosomes and neutrophil extracellular traps (NETs) such as:
• Oncology
• Coagulation
• Sepsis
• Auto-immune Diseases
• Neurodegenerative Diseases
• Inflammation
• NETopathies (neutrophil extracellular trap associated diseases)
• COVID-19

Assay Principle

The Nu.Q^® Discover H3.1 ELISA Research Use Only kit is a sandwich format colorimetric assay for research use only (RUO). The sample is added with the assay buffer to the well coated with anti-H3.1 antibody. After a first incubation followed by a washing step, the detection antibody coupled to HRP is added. After a second incubation and washing step, the TMB substrate is added. The absorbance is read after addition of stop solution. The signal obtained is proportional to the concentration of H3.1 nucleosome present in the sample. This colorimetric assay is provided in a 96-stripwell format, enabling high and low throughput analysis.

Convenience:
• cf-nucleosome quantification technology run manually on ELISA sandwich immunoassay platform.
• The procedure is streamlined and time-efficient, allowing completion in just 2.5 hours.
• No assay development required, assay ready to run.
• The assay’s strip microplate design offers adaptability for both manual processing and high-throughput automation

Sensitivity & Specificity:
• Low sample volumes. Validated for EDTA plasma, cell culture extract, and supernatant; contact us for guidance on other biofluids.
• Targets the H3.1 variant associated with cell proliferation, relevant in cancer and other diseases.
• Detection antibody recognizes a nucleosome specific epitope ensuring detection of only intact nucleosomes.
• Typical reproducibility:
– Precision for Nu.Q^® H3.1 intra-run less than 15%CV.
– Precision for Nu.Q^® H3.1 inter-run less than 20%CV.
• The reportable range of the Nu.Q° Discover H3.1 ELISA Assay – RUO is 22.7 ng/mL to 650 ng/mL.

Quality:
• Assay developed based on CLSI guidelines.
• Expert support for your research need

All reagents are exclusively for Research Use Only – not for use in diagnostic procedures

Material required but not provided

• Deionized water.
• Adjustable or preset single-channel pipettes able to dispense 20μL.
• Multi-channel pipettes able to dispense 50μL to 200μL.
• Disposable pipette tips.
• Disposable reagent reservoirs.
• Vortex mixer.
• Volumetric cylinders.
• Container for biohazardous waste.
• Microplate incubator with orbital shaking facility (shaker at 700 rpm).
• Microplate reader equipped with 450nm filter
• Absorbent paper.
• Light-tight plate cover.

REMARK: For best assay efficiency, we recommend the use of qualified pipettes, shaker, and reader.

Standardized

Analytically validated quantitative assays calibrated in ng/ml using recombinant nucleosome standards for use in EDTA plasma. The Nu.Q^® H3.1 ELISA Assay was developed based on CLSI guidelines.

Quantification

Recombinant nucleosomes enable absolute quantification of levels of circulating H3.1 nucleosomes, ideal to measure apoptosis or release of histone complexes in research.

Detection Range

The reportable range of the Nu.Q^® Discover H3.1 ELISA Assay is 22.7 ng/mL to 650 ng/mL.

Analytical Performance

Sensitivity: The Lower Limit of Detection is 9.1ng/mL.

Specificity: Nucleosome epitope specific antibody enables detection of only intact nucleosomes

Interferences

The following potential interfering substances were tested and have shown not to interfere on the assay when present below the concentrations defined in the following table:

Species Cross-Reactivity

Nucleosome structure is conserved across species. H3.1 has been evaluated in samples taken from human and multiple animal models including mouse, dogs, cats, horses, rats, and pigs.

Please use in conjunction with the instructions for use.