ELISA Kit for Deoxyribonuclease I (DNASE1) Homo sapiens (Human) Sandwich ELISA

DNaseI; DNL1; DRNI; DNase-I; Dornase alfa

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  • ELISA Kit for Deoxyribonuclease I (DNASE1) Packages (Simulation)
  • ELISA Kit for Deoxyribonuclease I (DNASE1) Packages (Simulation)
  • ELISA Kit for Deoxyribonuclease I (DNASE1) Results demonstration
  • SEB127Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Deoxyribonuclease I (DNASE1) and the recovery rates were calculated by comparing the measured value to the expected amount of Deoxyribonuclease I (DNASE1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 87-96 90
EDTA plasma(n=5) 96-103 99
heparin plasma(n=5) 96-103 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Deoxyribonuclease I (DNASE1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Deoxyribonuclease I (DNASE1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Deoxyribonuclease I (DNASE1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 98-105% 79-105% 80-102% 98-105%
EDTA plasma(n=5) 96-103% 95-103% 83-103% 79-101%
heparin plasma(n=5) 93-104% 86-99% 89-96% 99-105%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Deoxyribonuclease I (DNASE1)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Deoxyribonuclease I (DNASE1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Deoxyribonuclease I (DNASE1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Deoxyribonuclease I (DNASE1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Deoxyribonuclease I (DNASE1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Low Circulating Levels of Mitochondrial and High Levels of Nuclear DNA Predict Mortality in Chronic Heart FailurePubmed:27349571
  • Inhibition of nuclease activity by a splice-switching oligonucleotide targeting deoxyribonuclease 1 mRNA prevents apoptosis progression and prolong viability of …Pubmed: 32315661
  • Biomarkers of neutrophil extracellular traps (NETs) and nitric oxide-(NO)-dependent oxidative stress in women who miscarriedPubmed: 32753622
  • DNase 1 Protects From Increased Thrombin Generation and Venous Thrombosis During Aging: Cross‐Sectional Study in Mice and HumansPubmed:35023342

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