Tissue plasminogen activator (tPA) is a serine protease that converts plasminogen to plasmin in the blood fibrinolytic system. It also plays an important role in the nervous system, including the processes of neuronal migration, neurite outgrowth, and neuronal plasticity. tPA has been suggested to have a role in several neuropathological conditions such as cerebral ischemia, seizures, and demyelinating diseases. The sensitive quantitative measurement of total mouse tPA antigen in plasma and other biological fluid samples is easily performed with this 96 well strip format ELISA kit. The level of tPA antigen in normal plasma was found to be 9.4 ng/ml in NSA/CF-1, 2.4 ng/ml in C57BL6, and 0.4 ng/ml in CD-1 mouse strains. The assay measures total tPA in the 0.1-50 ng/ml range. Samples giving mouse tPA levels above 50 ng/ml should be diluted in blocking buffer before use. Mouse tPA will bind to the capture antibody coated on the microtiter plate. Free and complexed tPA will be detected by the assay. After appropriate washing steps, anti-mouse tPA primary antibody binds to the captured protein. Excess primary antibody is washed away and bound antibody is reacted with the secondary antibody conjugated to HRP. Following an additional washing step, TMB substrate is used for color development at 450 nm. Color development is proportional to the concentration of mouse tPA in the samples. A standard calibration curve is prepared in blocking buffer using dilutions of purified mouse tPA and is measured along with the test samples. All reagents and standards are provided in these ELISA kits.
Suggested additional reagents: 10X Wash Buffer, TMB Substrate, tPA Antigen Capture Plate, Secondary Antibody