Description
Plasminogen activator inhibitor type 1 (PAI-1) is involved in the regulation of the blood fibrinolytic system. Increased plasma levels of PAI-1 are implicated in the impairment of fibrinolytic function and may be associated with thrombotic diseases. Levels of PAI-1 increase with age and are elevated in conditions such as normal pregnancy and sepsis. The sensitive quantitative measurement of functionally active canine (dog) PAI-1 in plasma samples is easily performed with this 96 well strip format ELISA kit. The assay measures active PAI-1 in the 0.2-100 ng/ml range. Samples giving mouse PAI-1 levels above 100 ng/ml should be diluted in blocking buffer before use. Normal plasma should be applied directly to the plate for best results. It is important to ensure a platelet free preparation of plasma as platelets can release PAI-1. Functionally active PAI-1 reacts with urokinase coated onto a micro titer plate. Latent or complexed PAI-1 will not bind to the plate and will not be detected by the assay. After appropriate washing steps, anti-dog PAI-1 primary antibody binds to the captured enzyme. Excess 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 PAI-1 in the samples. A standard calibration curve is prepared using dilutions of purified PAI-1 and is measured along with the test samples. All reagents and standards are provided in these ELISA kits