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resDetect™ DNase Activity Assay Kit (Fluorescence)

For research use only.
    Product Specifications
    Assay TypeFRET
    AnalyteDNase I
    Format96T/480T
    Regulatory StatusRUO
    Sensitivity3.9×10⁻⁵ U
    Standard Curve Range0.0025 U-3.9×10⁻⁵ U
    Assay Time30 min
    Suitable Sample TypeFor the quantitative determination of DNase I in the environment, some biological materials, common molecular biological reagents such as reaction buffers.
    Sample volume10 μL & 80 μL
  • Product Overview
    resDetect™ DNase Activity Assay Kit (Fluorescence) is based on the FRET method and is used to measure the presence of DNase in buffers, reagents, and other components within 30 minutes. It is designed to provide a rapid, highly sensitive, and quantitative universal detection tool for identifying DNase contamination that may affect experiments. Moreover, the DNase and RNase Activity Assay Kit (Fluorescence) has been designed to work seamlessly together for the simultaneous quantitative detection of DNases and RNases in a single sample.
  • Reconstitution
    Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
  • Storage
    1. Unopened kit should be stored at -25~-15℃ upon receiving.

    2. Find the expiration date on the outside packaging and do not use reagents past their expiration date.

    3. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

    Materials Provided
    ItemsSize (96tests)Size(480tests)
    DNase Substrate2 nmol10 nmol
    10X Reaction Buffer for DNase10 mL10 mL
    DNase I (1U/μL)20 μL50 μL
    TE Buffer (pH 8.0)1.5 mL6 mL
    Nuclease-free Water10 mL50 mL
  • Assay Principles
    The DNase Activity Assay Kit (Fluorescence) is based on a fluorophore-labeled DNase substrate. When the sample does not contain DNase activity, the substrate is stable and does not produce a fluorescent signal; when the sample contains DNase activity, the substrate is degraded, resulting in a gradual enhanced fluorescence signal, the rate of increase in fluorescence signal is positively correlated with the number and activity of enzymes. Use a fluorescence microplate reader to measure at the wavelength of ex/em = 535/565 nm to determine whether the sample is contaminated by DNase.

    Assay Principles

Bioactivity-Fluorescence Please refer to DS document for the assay protocol.
 DNase I FLUORESCENCE

Add 90 μL of the working DNase Substrate solution to each 96-well plate, and add 10 μL of DNase I standards (0.00000390625-0.00025 U/μL*10μL/well = 0.0000390625-0.0025U/well), incubate the plate in the fluorometer (BMG CLARIOstar) collecting real-time data at 1 minute intervals for 30 minutes at 37°C using the settings described in this section. The DNase Activity Assay can be evaluated in rigorous kinetic terms using real-time data.

 DNase I FLUORESCENCE

This assay kit employs a standard detection of DNase I. Add 90 μL working DNase Substrate solution to each 96-well plate, and add 10 μL DNase I standards ((0.00000390625-0.00025 U/μL*10μL/well = 0.0000390625-0.0025U/well), incubate for 30 minutes at 37°C. Then measure the fluorescence using the settings described in this section in a fluorometer (BMG CLARIOstar). Take RFU of standards as the ordinate and DNase concentration as the abscissa. Four parameters logistic are used to draw the standard curve (QC tested).

Validation
Intra-Assay Statistics

Eight replicates of each of seven samples containing different concentrations of DNase were tested in one assay, Intra-Assay Precision CV<10%.

 DNase I INTRA-ASSAY STATISTICS
Inter-Assay Statistics

Eight replicates of each of seven samples containing different concentrations of DNase were tested in eight independent assays, Inter-Assay Precision CV<15%.

 DNase I INTER-ASSAY STATISTICS
Stability

The probe is freeze-thawed 3 times, the kit performance meets the requirements, the sensitivity is not reduced, and the CV< 10%.

 DNase I STABILITY
Recovery

Three different concentrations of DNase was spiked into four different kinds of matrixes. The range of the recovery rate is 80%~120%.

 DNase I RECOVERY
Interference effect

DNase Activity Assay Kit (Fluorescence) (Cat. No. ASE-A002) and RNase Activity Assay Kit (Fluorescence) (Cat. No. ASE-A001) were used to detect nuclease residues in the same sample. No significant cross-reactivity or interference was observed.

 DNase I INTERFERENCE EFFECT
*Complete Your Research: RNase Activity Assay Kit (Fluorescence) (ACROBiosystems, cat#ASE-A001) is prepared.
  • Clinical and Translational Updates

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