Overview

  • Product name

    Mouse PAI1 ELISA Kit (SERPINE1)
    See all PAI1 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 6.7%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 14.2%
  • Sample type

    Cell culture supernatant, Serum, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    200 pg/ml
  • Range

    298 pg/ml - 5000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 109 90% - 121%
    Cell culture media 83 72% - 88%
    Cit plasma 104 96% - 111%
    serum free media 117 104% - 128%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    PAI1 (SERPINE1) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of PAI1 protein in mouse serum, plasma and cell culture supernatant.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Plasminogen activator inhibitor-1 (PAI1 or Serpin E1) is encoded by the Serpine1 gene. PAI1 is a serine protease inhibitor with a major role in the regulation of fibrinolysis. PAI1 is the principal inhibitor of tissue plasminogen activator (tPA) and urokinase (uPA), both of which are activators of plasminogen. PAI1 is a secreted protein produced by the endothelium as well as adipose tissue.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse PAI1 Capture Antibody 1 x 600µl
    10X Mouse PAI1 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    Mouse PAI1 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Serine protease inhibitor. This inhibitor acts as 'bait' for tissue plasminogen activator, urokinase, protein C and matriptase-3/TMPRSS7. Its rapid interaction with PLAT may function as a major control point in the regulation of fibrinolysis.
  • Tissue specificity

    Found in plasma and platelets and in endothelial, hepatoma and fibrosarcoma cells.
  • Involvement in disease

    Defects in SERPINE1 are the cause of plasminogen activator inhibitor-1 deficiency (PAI-1D) [MIM:613329]. It is a hematologic disorder characterized by increased bleeding after trauma, injury, or surgery. Affected females have menorrhagia. The bleeding defect is due to increased fibrinolysis of fibrin blood clots due to deficiency of plasminogen activator inhibitor-1, which inhibits tissue and urinary activators of plasminogen.
    Note=High concentrations of SERPINE1 seem to contribute to the development of venous but not arterial occlusions.
  • Sequence similarities

    Belongs to the serpin family.
  • Post-translational
    modifications

    Inactivated by proteolytic attack of the urokinase-type (u-PA) and the tissue-type (TPA), cleaving the 369-Arg-
    -Met-370 bond.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • Endothelial plasminogen activator inhibitor
    • PAI
    • PAI-1
    • PAI1
    • PAI1_HUMAN
    • PLANH1
    • Plasminogen activator inhibitor 1
    • Serine (or cysteine) proteinase inhibitor clade E (nexin plasminogen activator inhibitor type 1) member 1
    • Serpin E1
    • Serpin peptidase inhibitor clade E (nexin plasminogen activator inhibitor type 1) member 1
    • SERPINE1
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab197752 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of PAI1 were measured in duplicate and interpolated from the PAI1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PAI1 concentration was determined to be 18.98 ng/mL in mouse serum and 7.69 ng/mL in mouse plasma (citrate).

  • Background subtracted data values (mean +/- SD, n = 2) are graphed.

  • L929 cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. PAI1 was measured in 10-fold diluted cell culture supernatants of unstimulated and PMA/PHA stimulated L929 and cell culture media. Measured values were interpolated from the PAI1 Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed: 17.9 ng/mL unstimulated, 31.1 ng/mL stimulated, and undetectable in media.

Protocols

References

This product has been referenced in:

  • Li B  et al. MicroRNA-30b protects myocardial cell function in patients with acute myocardial ischemia by targeting plasminogen activator inhibitor-1. Exp Ther Med 15:5125-5132 (2018). Read more (PubMed: 29805539) »
  • Ma J  et al. Rivaroxaban attenuates thrombosis by targeting the NF-?B signaling pathway in a rat model of deep venous thrombus. Int J Mol Med 40:1869-1880 (2017). Read more (PubMed: 29039441) »
See all 2 Publications for this product

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