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Product Specifications
Assay TypeFRETAnalyteDNase IFormat96T/480TRegulatory StatusRUOSensitivity3.9×10⁻⁵ UStandard Curve Range0.0025 U-3.9×10⁻⁵ UAssay Time30 minSuitable 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 μLProduct 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 3 months from the date of opening.
Materials Provided
ItemsSize （96tests）Size（480tests）DNase Substrate2 nmol10 nmol10X Reaction Buffer for DNase10 mL10 mLDNase I (1U/μL)20 μL50 μLTE Buffer (pH 8.0)1.5 mL6 mLNuclease-free Water10 mL50 mLAssay 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.
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Performance Data
Bioactivity-Fluorescence
Please refer to DS document for the assay protocol.
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.
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).
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%.
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%.
Stability
The probe is freeze-thawed 3 times, the kit performance meets the requirements, the sensitivity is not reduced, and the CV< 10%.
Recovery
Three different concentrations of DNase was spiked into four different kinds of matrixes. The range of the recovery rate is 80%~120%.
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.
- *Complete Your Research: RNase Activity Assay Kit (Fluorescence) (ACROBiosystems, cat#ASE-A001) is prepared.
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Deoxyribonuclease 1
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