Myeloperoxidase (MPO) is a glycoprotein with a alpha2beta2 heteromultimer expressed in all cells of the myeloid linage. MPO is abundantly present in azurophilic granules of polymorphonuclear neutrophils. It is an important enzyme used during phagocytic lysis of engulfed foreign particles which takes part in the defense of the organism through production of hypochlorous acid (HOCl), a potent oxidant. MPO is rapidly released by activated polymorphonuclear neutrophils. Involvement of MPO has been described in numerous diseases such as atherosclerosis, lung cancer, Alzheimer’s disease and multiple sclerosis. Autoimmune antibodies to MPO are involved in Wegeners disease. Since the discovery of MPO deficiency, initially regarded as rare and restricted to patients suffering from severe infections, MPO has attracted more clinical attention.
The mouse MPO ELISA kit is to be used for the in vitro quantitative determination of mouse MPO in plasma, tissue homogenate and cell culture supernatant samples. Note that most reliable results are obtained with heparin plasma.
The mouse MPO ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours. The efficient format of a plate with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are incubated in microtiter wells coated with antibodies recognizing mouse MPO. Biotinylated tracer antibody will bind to captured mouse MPO. Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody. Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic acid. The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the mouse MPO standards (log). The mouse MPO concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
The linearity of the assay was determined by serially diluting a serum sample containing 49.62 ng/ml mouse MPO. The diluted samples were measured in the assay. The line obtained a slope of 1.03 and a correlation coefficient of 0.998.