Description The Immuchrom Pancreatic Elastase ELISA Kit is an immunoassay that enables easy, rapid and precise quantitative determination of pancreatic elastase in human stool samples. For research use only. Summary The pancreatic elastase belongs to the class of serine proteases and has a molecular weight of 26 kDa. It is a digestive enzyme that is produced in the pancreas of all vertebrates. The pancreatic elastase is produced in the pancreas as an inactive zymogen (proelastase) and activated in the small intestine by cleavage with trypsin. The elastase is excreted unmodified in the stool. The stool concentration of pancreatic elastase provides information about the performance of the pancreas. The ImmuChrom complete pancreatic elastase kit allows an easy, rapid and precise quantitative determination of pancreatic elastase in biological samples. The kit includes all reagents ready for use, except the washing buffer. Type Sandwich ELISA, HRP-labelled antibody Other names Chymotrypsin-like elastase family member 1, Pancreatic elastase-1, Pancreatic elastase 1, E1, pancreatic endopeptidase E, CELA1, ELA1, EC:3.4.21.36, UniProtKB# Q9UNI1. Principle of Method The pancreatic elastase-ELISA test determines human pancreatic elastase according to the “sandwich”-principle. The elastase in sample, standard and controls binds to antibodies, which are coated to the microtiter plate. After a washing step a peroxidase labeled detection antibody is added. A second washing step is followed by the addition of the substrate which is converted to a colored product by the peroxidase. The reaction is terminated by the addition of an acidic stop solution. The optical densities are read at 450 nm (against the reference wavelength 620 nm) in a microtiter plate reader. The pancreatic-elastase concentration can be calculated from the standard curve. Calibration: The test system is calibrated using a reference preparation of recombinant and purified human pancreatic elastase. Sample Types Stool Sample Volume Extraction in stick vials In a stool sample extraction vial mix 15 mg stool with 1.5 ml extraction buffer (EXT), then vortex it until the mixture is homogenous. Transfer the resulting slurry to a plastic vial and centrifuge it for 10 min at 3000 xg. The supernatant is diluted 1:50 in sample buffer (SAMPLEBUF). (5 µl supernatant + 245 µl sample buffer) 40 µl of the dilution are used in the test per well. Standard Curve Calibration Range: 24.4 - 660 µg/ml Limit of Detection 1.25 µg/ml For the determination the zero-standard was measured 20 times. The 3-fold standard deviation was added to the mean value of the optical density. The respective concentration was read from the standard curve. Intra-assay (Within-Run) C.V.
