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 total rat PAI-1 antigen in plasma samples is easily performed with this 96 well strip format ELISA kit. The concentration level of PAI-1 antigen in rat plasma was found to be 1.8 ng/ml. The assay measures rat PAI-1 in the 0.05-50 ng/ml range. Samples giving rat PAI-1 levels above 50 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. Rat PAI-1 will bind to the capture antibody coated on the microtiter plate. Free, latent, and complexed PAI-1 will be detected by the assay. After appropriate washing steps, biotin labeled anti-rat PAI-1 primary antibody binds to the captured protein. Excess primary antibody is washed away and bound antibody is reacted with peroxidase conjugated streptavidin. Following an additional washing step, TMB substrate is used for color development at 450 nm. Color development is proportional to the concentration of rat PAI-1 in the samples. A standard calibration curve is prepared in blocking buffer using dilutions of purified rat PAI-1 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, Rat PAI-1 Antigen Capture Plate, Secondary Antibody