Active human PAI-1 functional assay ELISA kit
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 plasma samples is easily performed with this 96 well strip format ELISA kit. The PAI-1 concentration of normal platelet-free plasma is 21 ng/ml, platelet-rich plasma is 283 ng/ml and serum is 270 ng/ml. 1 PAI-1 unit = 1.34 ng. The assay measures active PAI-1 in the 0.125-100 U/ml range. Samples giving human PAI-1 levels above 100 U/ml should be diluted in PAI-1 depleted plasma 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. Validated in citrate, EDTA, and heparin plasma. 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 PAI-1 depleted plasma using dilutions of purified PAI-1 and is measured along with the test samples. All reagents and standards are provided in these ELISA kits.The Human PAI-1 standard provided in this kit is calibrated against the WHO International Standard for Human PAI-1 (NIBSC Code 92/654).This assay uses an exclusive enzyme capture technology to only detect functionally active protein.
The standard curve for this kit is generated using included PAI-1 depleted human plasma and is best suited for measurement of PAI-1 in plasma or serum. This kit is also available with a standard curve in BSA/TBS for researchers measuring PAI-1 in culture media or tissue extracts under catalog number HPAIKT-NP.
Suggested additional reagents: 10X Wash Buffer, TMB Substrate, uPA Plate, Secondary Antibody
Gene ID: 5054
Swiss-Prot/UniProt ID: P05121
View sample datasheet
View kit instruction sheet
View sample certificate of analysis
View material safety data sheet
Ask a question about this product
1. Richardson MA, Gupta A, O’brien LA et-al. Treatment of sepsis-induced acquired protein C deficiency reverses Angiotensin-converting enzyme-2 inhibition and decreases pulmonary inflammatory response. J. Pharmacol. Exp. Ther. 2008;325 (1): 17-26. doi:10.1124/jpet.107.130609 – Pubmed citation
2. Sereda MJ, Bradding P, Vial C. Adenosine potentiates human lung mast cell tissue plasminogen activator activity. J. Immunol. 2011;186 (2): 1209-17. doi:10.4049/jimmunol.1001563 – Pubmed citation
3. Leong HS, Podor TJ, Manocha B et-al. Validation of flow cytometric detection of platelet microparticles and liposomes by atomic force microscopy. J. Thromb. Haemost. 2011;9 (12): 2466-76. doi:10.1111/j.1538-7836.2011.04528.x – Pubmed citation
4. Wilhelmsen K, Mesa KR, Lucero J et-al. ERK5 protein promotes, whereas MEK1 protein differentially regulates, the Toll-like receptor 2 protein-dependent activation of human endothelial cells and monocytes. J. Biol. Chem. 2012;287 (32): 26478-94. doi:10.1074/jbc.M112.359489 – Free text at pubmed – Pubmed citation
5. Kupchak BR, Creighton BC, Aristizabal JC et-al. Beneficial effects of habitual resistance exercise training on coagulation and fibrinolytic responses. Thromb. Res. 2013;131 (6): e227-34. doi:10.1016/j.thromres.2013.02.014 – Pubmed citation
6. Komissarov AA, Florova G, Azghani A et-al. Active ?-macroglobulin is a reservoir for urokinase after fibrinolytic therapy in rabbits with tetracycline-induced pleural injury and in human pleural fluids. Am. J. Physiol. Lung Cell Mol. Physiol. 2013;305 (10): L682-92. doi:10.1152/ajplung.00102.2013 – Free text at pubmed – Pubmed citation
7. Bondu V, Schrader R, Gawinowicz MA, et al. Elevated cytokines, thrombin and PAI-1 in severe HCPS patients due to Sin Nombre virus. Viruses. 2015;7(2):559-89. Link to article
8. Mashiko S, Kitatani K, Toyoshima M, et al. Inhibition of plasminogen activator inhibitor-1 is a potential therapeutic strategy in ovarian cancer. Cancer Biol Ther. 2015;16(2):253-60 Link to article
9. Balc? ekmekçi Ö, Ekmekçi H, Güngör Z, et al. Evaluation of Lp-PLA2 mass, vitronectin and PAI-1 activity levels in patients with preeclampsia. Arch Gynecol Obstet. 2015;292(1):53-8. Link to article
10. Zhang GY, Wu LC, Dai T, et al. NADPH oxidase-2 is a key regulator of human dermal fibroblasts: a potential therapeutic strategy for the treatment of skin fibrosis. Exp Dermatol. 2014;23(9):639-44. Link to article
11. Ohkura N, Oiwa H, Ohnishi K, Taniguchi M, Baba K, Atsumi G. Inhibition of plasminogen activator inhibitor-1 release from human endothelial cells by Angelica keiskei Koidzumi (Ashitaba) chalcones is structure-dependent. J Intercult Ethnopharmacol. 2015;4(4):355-7.
12. Luo M, Li R, Ren M, et al. Hyperglycaemia-induced reciprocal changes in miR-30c and PAI-1 expression in platelets. Sci Rep. 2016;6:36687. Link to article.
13. Chang ML, Lin YS, Pao LH, Huang HC, Chiu CT. Link between plasminogen activator inhibitor-1 and cardiovascular risk in chronic hepatitis C after viral clearance. Sci Rep. 2017;7:42503. Link to article
14. Mcmahon GA, Petitclerc E, Stefansson S, et al. Plasminogen activator inhibitor-1 regulates tumor growth and angiogenesis. J Biol Chem. 2001;276(36):33964-8. Link to article
15. Roberson PA, Chase JD, Bigman MB, Saunders MJ, Luden ND, Womack CJ. Time of Day, but not Sleep Restriction, Affects Markers of Hemostasis Following Heavy Exercise. Appl Physiol Nutr Metab. 2018; Link to article