Description
Background/Introduction: 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.
Principle of the Assay: This assay employs a two-site sandwich ELISA to quantitative PR3 in Human serum, plasma. An antibody specific for PR3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any PR3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin - conjugated antibody specific for PR3 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 bound in the initial step. The color development is stopped and the intensity of the color is measured