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Nu.Q™ Total

determine total levels of circulating cell-free nucleosomes

Active Motif’s Nu.Q™ Total Assay Kit is designed for the detection of global levels of cell-free circulating nucleosomes (cf-nucleosomes) in human serum in a high-throughput format. For added convenience and a more quantitative interpretation of results, the Nu.Q™ Total Assay Kit also include a recombinant nucleosome protein for use as a reference standard curve. For complete details, click the Nu.Q™ Method tab below.

Why use Nu.Q™ Total?

  • Sensitivity: Detect circulating nucleosomes in as little as 10 µl of serum
  • Specificity: Nucleosome epitope specific antibody enables detection of only intact nucleosomes
  • Quantification: Recombinant nucleosomes enable relative quantification of total levels of circulating nucleosomes
  • Convenience: Colorimetric assay in a simple 96-stripwell format for high and low throughput
  • Fast: Results can be obtained in 5 hours

Video demonstrating the Nu.Q™ assay technology.

 

Active Motif's Nu.Q™ Total Assay Kit is powered by VolitionRx.

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Name Format Cat No. Price  
Nu.Q™ Total Assay Kit 1 x 96 rxns 53510 $545 Buy Now

Nu.Q™ Total Assay

Nucleosomes are repeating subunits of DNA and histone proteins that constitute human chromatin. Increased levels of nucleosomes in the blood have been associated with a number of pathological conditions and likely constitutes a major proportion of the cell free DNA (cfDNA) found in the bloodstream. Thus, quantification of the levels of circulating nucleosomes in serum gives insight into the progression of many conditions. The Nu.Q™ Total Assay enables high-throughput screening of cell-free circulating nucleosomes, regardless of histone modification states. This assay is able to detect total levels of circulating nucleosomes, from as little as 10 µl of human serum. (Figure 1).

Nu.Q™ Total Assay.
 
Figure 1: Total circulating cell-free nucleosome detection.

The Nu.Q™ Total Assay Kit was used to assay human serum samples. The provided Positive Control was assayed from 0.039 - 2.5 µg/ml as a reference standard curve. Data shown are the results from wells assayed in duplicate with a 20 minute developing time. These results are provided for demonstration only.

The Nu.Q™ Assay Method

The Nu.Q™ Total Assay Kit is a sandwich ELISA designed for the detection of global levels of cell-free circulating nucleosomes (cf-nucleosomes) in human serum. Its high-throughput format enables a large number of samples to be screened simultaneously. The assay utilizes a histone H3 monoclonal antibody to capture nucleosomes from your serum samples. Subsequently a biotinlylated recombinant antibody specific to a common nucleosome epitope is used for detection to assure that only intact nucleosomes are quantified in the assay. Streptavidin conjugated to horseradish peroxidase (SA-HRP) and developing solutions provide a sensitive colorimetric readout that is easily quantified by spectrophotometry. The assay is able to detect total levels of circulating nucleosomes, regardless of the modification state, from as little as 10 µl of human serum.

Included is a recombinant nucleosome made using Active Motif's patented protein synthesis technology. This protein can be used to build a reference standard curve to determine the total amount of circulating nucleosomes in your samples.

Flow Chart of Nu.Q™ Assay Method

Flow chart of Nu.Q™ Assay.
 
Figure 1: Flow chart of the Nu.Q™ Assay Method.

The Nu.Q™ Assay Method utilizes a histone H3 antibody to capture nucleosomes from samples and a nucleosome epitope specific antibody for detection. Streptavidin conjugated to horseradish peroxidase and developing solutions provide a colorimetric readout that is quantified by spectrophotometry. (Click image to enlarge.)

Contents & Storage

Nu.Q Total Assay Kits include a 96-stripwell Total Nucleosome Capture Plate, Total Detection Antibody, Detection Diluent, Streptavidin-HRP Conjugate, Assay Buffer, 10X Wash Buffer, Substrate Solution, Stop Solution, Total Recombinant Nucleosome Positive Control, Control Diluent, Stop Solution and plate sealer. Storage conditions vary from room temperature to -80°C, see manual for details. All reagents are guaranteed stable for 6 months when stored properly.