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 human PAI-1 in non-plasma samples including cell culture media or tissue extracts is easily performed with this 96 well strip format ELISA kit. The assay measures active PAI-1 in the 0.125-100 ng/ml range. Samples giving human PAI-1 levels above 100 ng/ml should be diluted in blocking buffer before use. 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. Vitronectin does not interfere with the detection of active PAI-1. This kit does not cross react with mouse PAI-1. After appropriate washing steps, anti human PAI-1 primary antibody binds to the PAI-1. 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 in blocking buffer using dilutions of purified PAI-1 and is measured along with the test samples. All reagents and standards are provided in these ELISA kits.
This assay uses an exclusive enzyme capture technology to only detect functionally active protein.
This kit is also available with a standard curve in PAI-1 depleted human plasma for researchers measuring PAI-1 in plasma or serum samples under catalog number HPAIKT.