PMN ELASTASE ELISA KIT
INTENDED USE
The PMN Elastase ELISA Kit is intended for the quantitative determination of PMN Elastasein serum, seminal plasma and stool. It is for in vitro diagnostic use only.
INTRODUCTION
PMN Elastase from human polymorphnuclear granulocytes is a glycoprotein of 30 kDa which belongs to the group of serine proteases. Active PMN Elastase is released from azurophil granula of neutrophil granulocytes after irritation or disintegration. The determination of the PMN Elastase in stool is used to record inflammatory reactions in which neutrophils are involved. Especially in Crohn´s disease the inflammatory process goes hand in hand with an increased phagocytic activity and the biological decay of the phagocytic cells, thus, leading to an increased release of PMN Elastase and other lysosomal enzymes.
PRINCIPLE OF THE TEST
PMN Elastase ELISA Kit
The PMN Elastase ELISA Kit is intended for the quantitative determination of PMN-Elastasein serum, seminal plasma and stool. In a first incubation step, PMN Elastase in the sample is bound to polyclonal sheep-anti-PMN-Elastase antibodies (in excess), which are immobilized on the surface of the microtiter wells (PLATE). To remove all unbound substances, a washing step is carried out. In a second incubation step, a monoclonal mouse-anti-PMN-Elastase antibody (AB) is added. This antibody is able to detect both the free and the complexed form with the specific inhibitor (α1-Proteinase inhibitor = α1-Antitrypsin). The quantification of the bound PMN-Elastase is carried out by adding an antimouse peroxidase-labeled conjugate (CONJ). Finally, the PMN-Elastaseantigen-antibody-complex is incubated with the peroxidase substrate, tetramethylbenzidine (SUB). An acidic stop solution (STOP) is then added to terminate the reaction. The color changes from blue to yellow. The intensity of the yellow color is directly proportional to the concentration of PMN Elastase in the sample. Samples are quantified by referring their optical density to a Lot-dependant master calibration curve and the use of a Calibrator that is run with each test. The combination of two specific antibodies in the PMN Elastase ELISA drastically reduces the possibility of false results and offers a reliable diagnostic system to the user.
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