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Scientific Background
PRTN3 is a serine protease enzyme expressed mainly in neutrophil granulocytes. Its exact role in the function of the neutrophil is unknown but in human neutrophils proteinase 3 contributes to the proteolytic generation of antimicrobial peptides. It is also the epitope of anti-neutrophil cytoplasmic antibodies (ANCAs) of the cANCA (cytoplasmic subtype) class, a type of antibody frequently found in the disease Wegener's granulomatosis.Proteinase K is commonly used in molecular biology to digest protein and remove contamination from preparations of nucleic acid. Addition of proteinase K to nucleic acid preparations rapidly inactivates nucleases that might otherwise degrade the DNA or RNA during purification.
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate PR3-ANCA in samples. An antibody specific for PR3-ANCA has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPR3-ANCA present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PR3-ANCA is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of PR3-ANCA bound in the initial step. The color development is stopped and the intensity of the color is measured.
Performance Data
| Assay Type | Sandwich ELISA |
|---|---|
| Detection Range | 0.156-10 ng/mL |
| Sensitivity | 0.061 ng/mL |
| Species Reactivity | Human (Homo sapiens) |
| Stability | The stability of ELISA 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. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C). |
| Manufacturer No. | AE63015HU |
Assay Workflow
The kit includes the components required for standard-curve quantification (see the kit manual for the component table and validated sample types). Absorbance is read on a standard microplate reader and concentrations are interpolated from the standard curve.
Safety & Handling
For research use only (RUO). Not for use in diagnostic or therapeutic procedures. Handle all samples and reagents using standard laboratory safety practices.
Related Products
Explore additional Human ELISA kits from Wuhan Abebio Science Co Ltd, or contact us for related assays for PR3-ANCA research.
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