Tissue plasminogen activator (tPA) is a serine protease that catalyzes the activation of plasminogen to plasmin. Clinical studies have indicated that high tPA levels may increase the risk for thrombosis, whereas decreased levels may cause neuronal plasticity and degeneration. The sensitive quantitative measurement of total human tPA antigen in plasma, serum, urine, cell culture media, or tissue extract samples is easily performed with this 96 well strip format ELISA kit. The basal level of tPA in healthy males and females, age 25-34 years, were found to be 5.5 ng/ml and 4.0 ng/ml respectively. The assay measures human tPA in the 0.02-10 ng/ml range. Samples giving human tPA levels above 10 ng/ml should be diluted in blocking buffer before use. Human tPA will bind to the affinity purified capture antibody coated on the microtiter plate. Free and complexed tPA will be detected by the assay. This kit cross reacts with mouse, rat, and pig tPA and does not cross react with rabbit, cyno monkey, rhesus monkey, guinea pig, dog, and sheep tPA. After appropriate washing steps, monoclonal anti-human tPA primary antibody binds to the captured protein. Excess primary antibody is washed away and bound antibody is reacted with peroxidase conjugated secondary antibody. Following an additional washing step, TMB substrate is used for color development at 450 nm. Color development is proportional to the concentration of human tPA in the samples. A standard calibration curve is prepared in blocking buffer using dilutions of purified human 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