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Human Serpin E1/PAI-1 DuoSet ELISA

Catalog #: DY1786
18 Citations 2 Reviews Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY1786
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human Serpin E1 / PAI-1 ELISA Standard Curve
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Product Details
Procedure
Citations (18)
FAQs
Supplemental Products
Reviews (2)
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human Serpin E1/PAI-1 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
0.3 - 20 ng/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Serpin E1/PAI-1. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

Normal Goat Serum: (Catalog # DY005)

 

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Normal Goat Serum: (Catalog # DY005)

 

Scientific Data

N/A Human Serpin E1 / PAI-1 ELISA Standard Curve View Larger

Human Serpin E1 / PAI-1 ELISA Standard Curve

Product Datasheets

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Product Datasheet
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SDS

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent with NGS, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human Serpin E1/PAI-1 DuoSet ELISA

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.

18 Citations: Showing 1 - 10
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  1. Dupuytren's Disease Is Mediated by Insufficient TGF-?1 Release and Degradation
    Authors: Oezel, L;Wohltmann, M;Gondorf, N;Wille, J;Güven, I;Windolf, J;Thelen, S;Jaekel, C;Grotheer, V;
    International journal of molecular sciences
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Evaluating circulating soluble markers of endothelial dysfunction and risk factors associated with PE: A multicentre longitudinal case control study in northern Ghana
    Authors: Ahenkorah, B;Sakyi, SA;Fondjo, LA;Helegbe, G;Owiredu, EW;Der, EM;Amoah, LE;Kusi, KA;Obiri, D;Amoani, B;Bimpong, S;Ofosu, W;Obirikorang, C;Odame, E;Larbie, C;Arthur-Johnson, P;Quaye, I;Ametefe, EM;Okai, BK;Anormah, R;Akorli, E;Simono Charadan, AM;Aboubacar, RM;Amesewu, E;Gyan, B;
    Heliyon
    Species: Human
    Sample Types: Serum
  3. Impaired Organokine Regulation in Non-Diabetic Obese Subjects: Halfway to the Cardiometabolic Danger Zone
    Authors: H L?rincz, B Ratku, S Csiha, I Seres, Z Szabó, G Paragh, M Harangi, S Somodi
    International Journal of Molecular Sciences, 2023-02-18;24(4):.
    Species: Human
    Sample Types: Plasma
  4. Obesity promotes radioresistance through SERPINE1-mediated aggressiveness and DNA repair of triple-negative breast cancer
    Authors: YH Su, YZ Wu, DK Ann, JL Chen, CY Kuo
    Cell Death & Disease, 2023-01-21;14(1):53.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Biomarkers for anti-vascular endothelial growth factor drugs
    Authors: S Kuriyama, T Yamada, A Matsuda, G Takahashi, T Iwai, K Takeda, K Ueda, T Miyasaka, Y Yokoyama, S Shinji, H Sonoda, R Ohta, K Yonaga, S Kanaka, H Yoshida
    Oncology Letters, 2022-11-07;24(6):463.
    Species: Human
    Sample Types: Plasma
  6. Mast Cell Proteases Promote Diverse Effects on the Plasminogen Activation System and Wound Healing in A549 Alveolar Epithelial Cells
    Authors: S Mogren, F Berlin, L Eskilsson, N Van Der Bu, E Tufvesson, CK Andersson
    Cells, 2022-09-18;11(18):.
    Species: Human
    Sample Types: BALF
  7. Clinical and biological markers for predicting ARDS and outcome in septic patients
    Authors: J Villar, R Herrán-Mon, E González-H, M Prieto-Gon, A Ambrós, A Rodríguez-, A Muriel-Bom, R Solano, C Cuenca-Rub, A Vidal, C Flores, JM González-M, MI García-Lao, Genetics o
    Scientific Reports, 2021-11-22;11(1):22702.
    Species: Human
    Sample Types: Serum
  8. Beneficial Effects of Mineralocorticoid Receptor Pathway Blockade against Endothelial Inflammation Induced by SARS-CoV-2 Spike Protein
    Authors: E Jover, L Matilla, M Garaikoetx, A Fernández-, P Muntendam, F Jaisser, P Rossignol, N López-Andr
    Biomedicines, 2021-06-03;9(6):.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Long intergenic non-coding RNAs regulate human lung fibroblast function: Implications for idiopathic pulmonary fibrosis
    Authors: MR Hadjichara, BT Roux, E Csomor, CA Feghali-Bo, LA Murray, DL Clarke, MA Lindsay
    Sci Rep, 2019-04-15;9(1):6020.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Biphasic activation of complement and fibrinolysis during the human nasal allergic response
    Authors: RS Thwaites, NC Gunawardan, V Broich, EH Mann, J Ahnström, GA Campbell, S Lindsley, N Singh, T Tunstall, DA Lane, PJ Openshaw, CM Hawrylowic, TT Hansel
    J. Allergy Clin. Immunol., 2018-02-07;0(0):.
    Species: Human
    Sample Types: Nasal Fluid
  11. Sustained low peritoneal effluent CCL18 levels are associated with preservation of peritoneal membrane function in peritoneal dialysis
    Authors: M Ossorio, MA Bajo, G Del Peso, V Martínez, M Fernández, MJ Castro, A Rodríguez-, R Madero, T Bellón, R Selgas
    PLoS ONE, 2017-04-17;12(4):e0175835.
    Species: Human
    Sample Types: Serum
  12. Human cytomegalovirus interleukin-10 enhances matrigel invasion of MDA-MB-231 breast cancer cells
    Authors: CA Valle Oseg, JV Spencer
    Cancer Cell Int, 2017-02-13;17(0):24.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Serum from Varicose Patients Induces Senescence-Related Dysfunction of Vascular Endothelium Generating Local and Systemic Proinflammatory Conditions
    Oxid Med Cell Longev, 2016-11-23;2016(0):2069290.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Characterization of In Vitro Engineered Human Adipose Tissues: Relevant Adipokine Secretion and Impact of TNF-alpha.
    Authors: Aubin K, Safoine M, Proulx M, Audet-Casgrain M, Côté J, Tetu F, Roy A, Fradette J
    PLoS ONE, 2015-09-14;10(9):e0137612.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Long-Term Expansion in Platelet Lysate Increases Growth of Peripheral Blood-Derived Endothelial-Colony Forming Cells and Their Growth Factor-Induced Sprouting Capacity.
    Authors: Tasev D, van Wijhe M, Weijers E, van Hinsbergh V, Koolwijk P
    PLoS ONE, 2015-06-15;10(6):e0129935.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Stimulated in vivo synthesis of plasminogen activator inhibitor-1 in human adipose tissue.
    Authors: Ekstrom M, Liska J, Eriksson P, Sverremark-Ekstrom E, Tornvall P
    Thromb Haemost, 2012-06-28;108(3):485-92.
    Species: Human
    Sample Types: Plasma
  17. Regulation of the endothelial plasminogen activator system by fluvastatin. Role of Rho family proteins, actin polymerisation and p38 MAP kinase.
    Authors: Dunoyer-Geindre S, Fish RJ, Kruithof EK
    Thromb. Haemost., 2010-12-21;105(3):461-72.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Prognostic impact of plasminogen activator inhibitor type 1 expression in bladder cancer.
    Authors: Becker M, Szarvas T, Wittschier M, vom Dorp F, Totsch M, Schmid KW, Rubben H, Ergun S
    Cancer, 2010-10-01;116(19):4502-12.
    Species: Human
    Sample Types: Plasma

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Reviews for Human Serpin E1/PAI-1 DuoSet ELISA

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Human Serpin E1/PAI-1 DuoSet ELISA
By Carmen Martin-Ruiz on 12/13/2021
Sample Tested: Serum

We used serum neat, diluted 1/5 and 1/10. It worked really well, very reproducible

Human Serpin E1/PAI-1 DuoSet ELISA
By Anonymous on 12/15/2020
Sample Tested: EDTA Plasma