Human PAI1 ELISA Kit (SERPINE1) (ab269373)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 16 pg/ml
- Range: 31.25 pg/ml - 2000 pg/ml
- Sample type: Cell culture supernatant, Cell Lysate, Cit plasma, EDTA Plasma, Hep Plasma, Serum
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human PAI1 ELISA Kit (SERPINE1)
See all PAI1 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 3 3.3% Inter-assay Sample n Mean SD CV% Serum 8 3.8% -
Sample type
Cell culture supernatant, Serum, Cell Lysate, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
16 pg/ml -
Range
31.25 pg/ml - 2000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 96 88% - 102% Serum 90 86% - 95% Cell Lysate 108 108% - 111% Hep Plasma 100 97% - 103% EDTA Plasma 96 94% - 97% Cit plasma 103 97% - 107% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human PAI1 ELISA Kit (SERPINE1) (ab269373) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of PAI1 (SERPINE1) protein in cell culture supernatant, cell lysate, edta plasma, hep plasma, serum, and cit plasma. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human PAI1 (SERPINE1) with 16 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
Assay Specificity: This kit recognizes both native and recombinant human PAI1 protein in serum, plasma, cell culture supernatant, and cell extract samples only. Tissue extract samples have not been tested with this kit.
Species Reactivity: This kit recognizes human PAI1 protein.
Other species reactivity was determined by measuring 1:20 (dilution) serum samples of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration in human serum assayed at the same dilution.
Monkey serum was 100% cross-reactive.
Reactivity
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Notes
Plasminogen activator inhibitor-1 (PAI-1) or SERPINE1is encoded by the SERPINE1 gene. The PAI-1 protein is a serine protease inhibitor (serpin) that functions as the principal inhibitor of tissue plasminogen activator (tPA) and urokinase (uPA). The inhibition of tPA and uPA leads to increased occurrence and persistence of blood clots to form.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human PAI1 (SERPINE1) Capture Antibody 1 x 600µl 10X Human PAI1 (SERPINE1) Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BI 1 x 6ml Human PAI1 (SERPINE1) 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
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Function
This inhibitor acts as 'bait' for tissue plasminogen activator, urokinase, and protein C. Its rapid interaction with TPA 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
modificationsInactivated 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
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Alternative names
- Clade E
- Endothelial plasminogen activator inhibitor
- Nexin
see all -
Database links
- Entrez Gene: 5054 Human
- Omim: 173360 Human
- SwissProt: P05121 Human
- Unigene: 414795 Human
- Unigene: 713079 Human
Images
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SimpleStep ELISA Protocol Diagram
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.
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Human PAI1 standard curve comparisonStandard Curve comparison between human PAI1 SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.
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Example of human PAI1 standard curve in Sample Diluent NS.The PAI1 standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Example of human PAI1 standard curve in Sample Diluent NS.The PAI1 standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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. Example of human PAI1 standard curve in 1X Cell Extraction Buffer PTR.The PAI1 standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Example of human PAI1 standard curve in 1X Cell Extraction Buffer PTR.The PAI1 standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Linearity of dilution.Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.
Native PAI1 was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
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Linearity of dilution.Native PAI1 was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
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Linearity of dilution.Native PAI1 was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
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Linearity of dilution.Native PAI1 was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in 1X Cell Extraction Buffer PTR.
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Interpolated concentrations of native PAI1 in human serum and plasma samples.The concentrations of PAI1 were measured in duplicates, interpolated from the PAI1 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 5%, plasma (citrate) 5%, plasma (EDTA) 10%, and plasma (heparin) 5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PAI1 concentration was determined to be 32.4 ng/mL in serum, 36.6 ng/mL in plasma (citrate), 11.1 ng/mL in plasma (EDTA), and 10.1 ng/mL in plasma (heparin).
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Interpolated concentrations of native PAI1 in human stimulated PBMC cell culture supernatant and unstimulated PBMC cell culture supernatant samples.The concentrations of PAI1 were measured in duplicates, interpolated from the PAI1 standard curves and corrected for sample dilution. Undiluted samples are as follows: stimulated PBMC cell culture supernatant 50% and unstimulated PBMC cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PAI1 concentration was determined to be 1,896 pg/mL in stimulated PBMC cell culture supernatant and 642 pg/mL in unstimulated PBMC cell culture supernatant.
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Interpolated concentrations of native PAI1 in HUVEC cell culture supernatant, and HepG2 cell culture supernatant samples.The concentrations of PAI1 were measured in duplicates, interpolated from the PAI1 standard curves and corrected for sample dilution. Undiluted samples are as follows: HUVEC cell culture supernatant 1:2,000 and HepG2 cell culture supernatant 1:400. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PAI1 concentration was determined to be 2,061 ng/mL in HUVEC cell culture supernatant and 739 ng/mL in HepG2 cell culture supernatant.
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Interpolated concentrations of native PAI1 in HUVEC cell extract and HepG2 cell extract samples based on a 500 µg/mL and 50 µg/mL extract load, respectively.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 677 pg/mL in HUVEC cell extract and 989 pg/mL in HepG2 cell extract.
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Serum from ten individual healthy human female donors was measured in duplicate.Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PAI1 concentration was determined to be 24 ng/mL with a range of 8 – 59 ng/mL.
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Assay sensitivity.The MDD was determined by calculating the mean of zero standard replicates and adding 2 standard deviations then extrapolating the corresponding concentration.
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Sandwich ELISA - Human PAI1 ELISA Kit (SERPINE1) (ab269373)To learn more about the advantages of recombinant antibodies see here.
