Human Serpin E1/PAI-1 Quantikine ELISA Kit

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Assay Procedure
Citations (24)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Cell Lysates (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Citrate Plasma (50 uL), CTAD Plasma (50 uL)
  • Sensitivity
    0.142 ng/mL
  • Assay Range
    0.3 - 20 ng/mL (Cell Culture Supernates, Cell Lysates, EDTA Plasma, Heparin Plasma, Citrate Plasma, CTAD Plasma)
  • Specificity
    Natural and recombinant human Serpin E1
  • Cross-reactivity
    Cross-reactivity observed with 1 or more available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Control Available
QC157 , Quantikine Immunoassay Control Set 778 Human Serpin E1/PAI-1 - Please Inquire
Product Summary
The Quantikine Human Serpin E1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Serpin E1 in cell culture supernates, cell lysates, and plasma. It contains Sf 21-expressed recombinant human Serpin E1 and has been shown to accurately quantitate the recombinant factor in its active and latent forms as well as in the vitronectin complexes. However, the kit does not detect recombinant human Serpin E1 in complexes with recombinant human uPA and recombinant human tPA. Results obtained using natural human Serpin E1 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Serpin E1.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates, Cell Lysates, EDTA Plasma, Heparin Plasma, Citrate Plasma, CTAD Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 2.6 6.5 12.6 2.3 6.4 13.1
Standard Deviation 0.12 0.52 0.98 0.2 0.4 0.8
CV% 4.4 8 7.8 9.5 6.4 6.1

Recovery

The recovery of Serpin E1 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 106 89-113
Citrate Plasma (n=4) 99 88-112
CTAD Plasma (n=4) 100 83-112
EDTA Plasma (n=4) 100 85-115
Heparin Plasma (n=4) 98 85-113
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Serpin E1 were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 Serpin E1/PAI-1 [HRP]
Product Datasheets

Certificate of Analysis Lookup

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Note: Certificate of Analysis not available for kit components.

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Serpin E1/PAI-1
The human serpin superfamily consists of at least 35 members that target not only serine proteases, but also selected cysteine proteases and non-protease proteins. Serpins bind the protease active site resulting in a major conformational rearrangement that traps the enzyme in a covalent acyl-enzyme intermediate. As protease inhibitors, serpins have an array of functions including regulating blood clotting, the complement pathway, extracellular matrix remodeling, and cell motility. They are also involved in activities that extend beyond their ability to inhibit proteases. For instance, they may also regulate blood pressure, angiogenesis, or act as storage/transport proteins.
  • Long Name:
    Plasminogen Activator Inhibitor
  • Entrez Gene IDs:
    5054 (Human); 18787 (Mouse)
  • Alternate Names:
    Endothelial plasminogen activator inhibitor; Nexin; PAI1; PAI-1; PAI1PAI-1; PAISerpin E1; PLANH1; PLANH1plasminogen activator inhibitor 1; serine (or cysteine) proteinase inhibitor, clade E (nexin, plasminogenactivator inhibitor type 1), member 1; Serpin E1; serpin peptidase inhibitor, clade E (nexin, plasminogen activator inhibitortype 1), member 1
Related Research Areas
Assay Procedure
Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

24 Citations: Showing 1 - 10
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Species
Sample Type
  1. Platelet Carbonic Anhydrase II, a Forgotten Enzyme, May Be Responsible for Aspirin Resistance
    Authors: M Jakubowski, J D?bski, E Szahidewic, A Turek-Jaku, J Gawry?, K Gawry?, R Skomro, A Derkacz, A Doroszko
    Oxid Med Cell Longev, 2017;2017(0):3132063.
    Species: Human
    Sample Type: Plasma
  2. Combined serum levels of multiple proteins in tPA-BDNF pathway may aid the diagnosis of five mental disorders
    Authors: S Chen, H Jiang, Y Liu, Z Hou, Y Yue, Y Zhang, F Zhao, Z Xu, Y Li, X Mou, L Li, T Wang, J Zhao, C Han, Y Sui, M Wang, Z Yang, Y Lu, Y Zhu, J Li, X Shen, F Sun, Q Chen, H Chen, Y Yuan
    Sci Rep, 2017;7(1):6871.
    Species: Human
    Sample Type: Serum
  3. Circadian misalignment increases cardiovascular disease risk factors in humans
    Proc. Natl. Acad. Sci. U.S.A., 2016;113(10):E1402-11.
    Species: Human
    Sample Type: Plasma
  4. MAP3K19 Is a Novel Regulator of TGF-? Signaling That Impacts Bleomycin-Induced Lung Injury and Pulmonary Fibrosis
    PLoS ONE, 2016;11(5):e0154874.
    Species: Human
    Sample Type: Cell Lysates
  5. Plasminogen Activator Inhibitor-1 in depression: Results from Animal and Clinical Studies
    Sci Rep, 2016;6(0):30464.
    Species: Human
    Sample Type: Serum
  6. Vitamin D insufficiency and subclinical atherosclerosis in non-diabetic males living with HIV.
    Authors: Portilla J, Moreno-Perez O, Serna-Candel C, Escoin C, Alfayate R, Reus S, Merino E, Boix V, Giner L, Sanchez-Paya J, Pico A
    J Int AIDS Soc, 2014;17(0):18945.
    Species: Human
    Sample Type: Serum
  7. Coinfection with human herpesvirus 8 is associated with persistent inflammation and immune activation in virologically suppressed HIV-infected patients.
    Authors: Masia M, Robledano C, Ortiz de la Tabla V, Antequera P, Lumbreras B, Hernandez I, Gutierrez F
    PLoS ONE, 2014;9(8):e105442.
    Species: Human
    Sample Type: Plasma
  8. Increased metabolite levels of glycolysis and pentose phosphate pathway in rabbit atherosclerotic arteries and hypoxic macrophage.
    Authors: Yamashita A, Zhao Y, Matsuura Y, Yamasaki K, Moriguchi-Goto S, Sugita C, Iwakiri T, Okuyama N, Koshimoto C, Kawai K, Tamaki N, Zhao S, Kuge Y, Asada Y
    PLoS ONE, 2014;9(1):e86426.
    Species: Human
    Sample Type: Cell Culture Supernates
  9. Ghrelin protects against renal damages induced by angiotensin-II via an antioxidative stress mechanism in mice.
    Authors: Fujimura K, Wakino S, Minakuchi H, Hasegawa K, Hosoya K, Komatsu M, Kaneko Y, Shinozuka K, Washida N, Kanda T, Tokuyama H, Hayashi K, Itoh H
    PLoS ONE, 2014;9(4):e94373.
    Species: Human
    Sample Type: Cell Culture Supernates
  10. Characterization of circulating and endothelial progenitor cells in patients with extreme-duration type 1 diabetes.
    Authors: Hernandez S, Gong J, Chen L, Wu I, Sun J, Keenan H, King G
    Diabetes Care, 2014;37(8):2193-201.
    Species: Human
    Sample Type: Plasma
  11. p-Cresol affects reactive oxygen species generation, cell cycle arrest, cytotoxicity and inflammation/atherosclerosis-related modulators production in endothelial cells and mononuclear cells.
    Authors: Chang M, Chang H, Chan C, Yeung S, Hsien H, Lin B, Yeh C, Tseng W, Tseng S, Jeng J
    PLoS ONE, 2014;9(12):e114446.
    Species: Human
    Sample Type: Whole Cells
  12. Staphylococcal nuclease domain containing-1 (SND1) promotes migration and invasion via angiotensin II type 1 receptor (AT1R) and TGFbeta signaling.
    Authors: Santhekadur P, Akiel M, Emdad L, Gredler R, Srivastava J, Rajasekaran D, Robertson C, Mukhopadhyay N, Fisher P, Sarkar D
    FEBS Open Bio, 2014;4(0):353-61.
    Species: Human
    Sample Type: Cell Culture Supernates
  13. Dipyridamole decreases inflammatory metalloproteinase-9 expression and release by human monocytes.
    Thromb Haemost, 2013;109(2):280-9.
    Species: Human
    Sample Type: Cell Culture Supernates
  14. Systemic biomarkers of inflammation and haemostasis in patients with chronic necrotizing pulmonary aspergillosis.
    BMC Infect Dis, 2012;12(0):144.
    Species: Human
    Sample Type: Plasma
  15. Recombinant GPI-anchored TIMP-1 stimulates growth and migration of peritoneal mesothelial cells.
    Authors: Djafarzadeh R, Sauter M, Notohamiprodjo S, Noessner E, Goyal P, Siess W, Wornle M, Ribeiro A, Himmelein S, Sitter T, Nelson PJ
    PLoS ONE, 2012;7(4):e33963.
    Species: Human
    Sample Type: Cell Lysates
  16. Exercise training improves sleep pattern and metabolic profile in elderly people in a time-dependent manner.
    Authors: Lira FS, Pimentel GD, Santos RV, Oyama LM, Damaso AR, Oller do Nascimento CM, Viana VA, Boscolo RA, Grassmann V, Santana MG, Esteves AM, Tufik S, de Mello MT
    Lipids Health Dis, 2011;10(0):113.
    Species: Human
    Sample Type: Plasma
  17. Endothelial differentiation of adipose-derived stem cells from elderly patients with cardiovascular disease.
    Authors: Zhang P, Moudgill N, Hager E, Tarola N, Dimatteo C, McIlhenny S, Tulenko T, DiMuzio PJ
    Stem Cells Dev., 2011;20(6):977-88.
    Species: Human
    Sample Type: Cell Culture Supernates
  18. Placenta growth factor (PlGF), a novel inducer of plasminogen activator inhibitor-1 (PAI-1) in sickle cell disease (SCD).
    Authors: Patel N, Sundaram N, Yang M
    J. Biol. Chem., 2010;285(22):16713-22.
    Species: Human
    Sample Type: Plasma
  19. Human CD4- 8- T cells are a distinctive immunoregulatory subset.
    Authors: Huang MC, Patel K, Taub DD, Longo DL, Goetzl EJ
    FASEB J., 2010;24(7):2558-66.
    Species: Human
    Sample Type: Cell Culture Supernates
  20. SPARC suppresses apoptosis of idiopathic pulmonary fibrosis fibroblasts through constitutive activation of beta-catenin.
    Authors: Chang W, Wei K, Jacobs SS, Upadhyay D, Weill D, Rosen GD
    J. Biol. Chem., 2010;285(11):8196-206.
    Species: Human
    Sample Type: Cell Culture Supernates
  21. Cyclic adenosine monophosphate induces plasminogen activator inhibitor-1 expression in human mast cells.
    Authors: Ma Z, Kwong KY, Tovar JP, Paek D
    Biochem. Biophys. Res. Commun., 2010;400(4):569-74.
    Species: Human
    Sample Type: Cell Culture Supernates
  22. Syndromic and non-syndromic aneurysms of the human ascending aorta share activation of the Smad2 pathway.
    Authors: Gomez D, Al Haj Zen A, Borges LF, Philippe M, Gutierrez PS, Jondeau G, Michel JB, Vranckx R
    J. Pathol., 2009;218(1):131-42.
    Species: Human
    Sample Type: Cell Culture Supernates
  23. Adiponectin regulates albuminuria and podocyte function in mice.
    Authors: Sharma K, Ramachandrarao S, Qiu G, Usui HK, Zhu Y, Dunn SR, Ouedraogo R, Hough K, McCue P, Chan L, Falkner B, Goldstein BJ
    J. Clin. Invest., 2008;118(5):1645-56.
    Species: Human
    Sample Type: Plasma
  24. Urokinase induces survival or pro-apoptotic signals in human mesangial cells depending on the apoptotic stimulus.
    Authors: Tkachuk N, Kiyan J, Tkachuk S, Kiyan R, Shushakova N, Haller H, Dumler I
    Biochem. J., 2008;415(2):265-73.
    Species: Human
    Sample Type: Cell Culture Supernates
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Quantikine Immunoassay Control Set 778 Human Serpin E1/PAI-1

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Cell Lysis Buffer 3 Concentrate (5X, 1 x 21 mL)

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 Human Serpin E1/PAI-1 Quantikine ELISA Kit DSE100
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Summary

Sample Tested cell culture supernatants

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Other Experimental Details used at 1:2 dilution for cell culture supernatants

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