Human PAI1+tPA ELISA Kit (ab269559)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 18 pg/ml
- Range: 78.13 pg/ml - 5000 pg/ml
- Sample type: Cell culture media, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Urine
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human PAI1+tPA ELISA Kit
See all PAI1 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 3 3.6% Inter-assay Sample n Mean SD CV% Serum 8 6.3% -
Sample type
Urine, Serum, Cell culture media, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
18 pg/ml -
Range
78.13 pg/ml - 5000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Urine 102 97% - 106% Serum 104 98% - 108% Cell culture media 85 82% - 87% Hep Plasma 88 84% - 92% EDTA Plasma 98 93% - 106% Cit plasma 99 96% - 106% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human PAI1+tPA ELISA Kit (ab269559) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of PAI1+tPA protein in edta plasma, hep plasma, serum, urine, cit plasma, and cell culture media. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human PAI1+tPA with 18 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.
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Notes
Tissue plasminogen activator (TPA or PLAT) is a serine protease that catalyzes the conversion of plasminogen to plasmin, which is integral to blood clot breakdown. Plasminogen activator inhibitor-1 (PAI1 or SERPINE1) is a serine protease inhibitor that blocks the function of TPA. PAI1 and TPA form a complex that is stable and non-covalent. The PAI1/TPA complex appears to be predictive of risk for myocardial infarction. This product uses a tPA specific capture antibody and a PAI1 specific detector antibody to specifically detect PAI1/TPA complexes in human biological samples. The reference protein is a non-covalent complex of PAI1 and TPA.
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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+tPA Capture Antibody 1 x 600µl 10X Human PAI1+tPA Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent 5BI 1 x 6ml Human PAI1+tPA Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml 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+tPA standard curve comparisonStandard Curve comparison between the original human MMP3 SimpleStep ELISA kit (ab92151) and current human MMP3 SimpleStep ELISA kit (ab269559).
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Example of human PAI1 + tPA standard curve in Sample Diluent NS.The PAI1 + tPA standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native PAI1 + tPA in human serum and plasma samples.The concentrations of PAI1 + tPA were measured in duplicates, interpolated from the PAI1 + tPA standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 25%, plasma (EDTA) 25%, and plasma (heparin) 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PAI1 + tPA concentration was determined to be 6.0 ng/mL in serum, 10.7 ng/mL in plasma (citrate), 4.3 ng/mL in plasma (EDTA) and 5.2 ng/mL in plasma (heparin).
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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 + tPA concentration was determined to be 7 ng/mL with a range of 3 – 12 ng/mL.
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Sandwich ELISA - Human PAI1+tPA ELISA Kit (ab269559)To learn more about the advantages of recombinant antibodies see here.