Human Serpin E1/PAI-1 DuoSet ELISA Summary
* 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
Product Datasheets
Preparation and Storage
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.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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.
20
Citations: Showing 1 - 10
Filter your results:
Filter by:
-
Baseline circulating biomarkers, their changes, and subsequent suicidal ideation and depression severity at 6 months: A prospective analysis in patients with mood disorders
Authors: Lengvenyte, A;Cognasse, F;Hamzeh-Cognasse, H;Sénèque, M;Strumila, R;Olié, E;Courtet, P;
Psychoneuroendocrinology
Species: Human
Sample Types: Plasma
-
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
-
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
-
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
-
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
-
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
-
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
-
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
-
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
-
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
-
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
-
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
-
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
-
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
-
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
-
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
-
Creating capillary networks within human engineered tissues: Impact of adipocytes and their secretory products
Authors: Kim Aubin, Caroline Vincent, Maryse Proulx, Dominique Mayrand, Julie Fradette
Acta Biomaterialia
Species: Human
Sample Types: Cell Culture Supernates
-
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
-
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
-
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
FAQs
No product specific FAQs exist for this product, however you may
View all ELISA FAQsReviews for Human Serpin E1/PAI-1 DuoSet ELISA
Average Rating: 4.5 (Based on 2 Reviews)
Have you used Human Serpin E1/PAI-1 DuoSet ELISA?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image
Filter by:
