Human PAI1 ELISA Kit (SERPINE1) (ab184863)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 21 pg/ml
- Range: 93.75 pg/ml - 6000 pg/ml
- Sample type: Cell culture supernatant, Cell Lysate, 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% HeLa extract 5 4.1% Inter-assay Sample n Mean SD CV% HeLa extract 3 9.1% -
Sample type
Cell culture supernatant, Serum, Plasma, Cell Lysate -
Assay type
Sandwich (quantitative) -
Sensitivity
21 pg/ml -
Range
93.75 pg/ml - 6000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 113 109% - 118% Plasma 115 115% - 116% Cell culture media 104 91% - 111% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human PAI1 ELISA kit has been re-developed. We have identified new recombinant monoclonal antibodies to provide improved performance and consistency. This version will be discontinued when inventory is depleted. The new version is available as ab269373.
Human PAI1 (SERPINE1) ELISA kit (ab184863) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of PAI1 protein in human serum, plasma, cell culture supernatants and cell extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate human PAI1 with 21 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 strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.Sensitivity:
Samples diluted in Sample Diluent 25BP - 48 pg/mL
Samples diluted in 1X Cell Extraction Buffer PTR - 21 pg/mLSPECIES REACTIVITY: This kit recognizes both native and recombinant human PAI1 protein in serum, plasma, cell culture supernatants and cell lysates.
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Notes
Plasminogen activator inhibitor-1 (PAI-1) is 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
Microplate
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human PAI1 Capture Antibody 1 x 600µl 10X Human PAI1 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 4BI 1 x 6ml Human PAI1 Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent 25BP 1 x 20ml 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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Other - Human PAI1 ELISA Kit (SERPINE1) (ab184863)
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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Figure 1Example of PAI1 standard curve using Sample Diluent 25BP. Background-subtracted data values (mean +/- SD) are graphed.
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Example of PAI1 standard prepared in Sample Diluent 25BP.Example of PAI1 standard prepared in Sample Diluent 25BP as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed
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Figure 2Example of PAI1 standard curve using 1X Cell Extraction Buffer PTR as a diluent. Background-subtracted data values (mean +/- SD) are graphed.
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Example of PAI1 standard prepared in 1X Cell Extraction Buffer PTR.Example of PAI1 standard prepared in 1X Cell Extraction Buffer PTR as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Titration of individual donor serum samples to within working range of the assay.Quantification of PAI interpolated from standard curve and multiplied by dilution factor.
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Figure 3Titration of cell culture supernatants diluted within the working range of the assay. A) Background subtracted data from duplicate measurements are plotted. B) Bar graph denotes quantification of PAI interpolated from standard curve and multiplied by dilution factor.
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Figure 4Titration of cell lysates diluted within the working range of the assay. Lysates made from HepG2 and HeLa cells were made according to Section 11. A) Background subtracted data from duplicate measurements of both lysates are plotted. HeLa lysates were found to be below the level of detection. B) Bar graph denotes quantification of PAI interpolated from standard curve and multiplied by dilution factor.
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Figure 5Titration of pooled normal human serum and plasma samples diluted within the working range of the assay. A) Background subtracted data from duplicate measurements are plotted. B) Bar graph denotes quantification PAI interpolated from standard curve and multiplied by dilution factor
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Assay sensitivity.The calculated minimal detectable dose (MDD) was determined by calculating the mean of zero standard replicates and adding 2 standard deviations then extrapolating the corresponding concentrations.
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Linearity of dilution.Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution
Linearity of dilution is determined based on interpolated values from the standard curve diluted in Sample Diluent 25BP. 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.
