Human TNF RI/TNFRSF1A 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 sTNF RI/TNFRSF1A. 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.
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)
Scientific Data
Product Datasheets
Preparation and Storage
Background: TNF RI/TNFRSF1A
TNF RI (TNF receptor 1; TNF R-p55/p60) is a transmembrane receptor for TNF-alpha and Lymphotoxin-alpha/TNF-beta. A soluble decoy form of the receptor is shed by ADAM-17/TACE in response to inflammatory stimulation. TNF RI signaling through NFkB is essential for the development of lymph node germinal centers and Peyer’s patches and for combating intracellular pathogens such as Listeria.
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, 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 TNF RI/TNFRSF1A 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.
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Citations: Showing 1 - 10
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Cognitive function is associated with performance in time up and go test and with leptin blood levels in community-dwelling older women
Authors: da Costa Teixeira, LA;Soares, LA;Lima, LP;Avelar, NCP;de Moura, JA;Leopoldino, AAO;Figueiredo, PHS;Parentoni, AN;Mendonça, VA;Lacerda, ACR;
Scientific reports
Species: Human
Sample Types: Plasma
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The strong inverse association between plasma concentrations of soluble tumor necrosis factor receptors type 1 with adiponectin/leptin ratio in older women
Authors: Augusto da Costa Teixeira, L;Rocha-Vieira, E;Aparecida Soares, L;Mota de Oliveira, F;Aparecida Oliveira Leopoldino, A;Netto Parentoni, A;Amaral Mendonça, V;Cristina Rodrigues Lacerda, A;
Cytokine
Species: Human
Sample Types: Plasma
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Association between biomarkers of redox status and cytokines with different patterns of habitual physical activity in eutrophic and overweight/obese preschoolers: multivariate analysis of a cross-sectional study
Authors: Viegas, ÂA;Santos, T;Nobre, JNP;Santos, JMD;Silva Lage, VKD;Fernandes, AC;Peixoto, MFD;Morais, RLS;Sartorio, A;Mendonça, VA;Lacerda, ACR;
BMC public health
Species: Human
Sample Types: Plasma
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Acute Kidney Injury-Induced Circulating TNFR1/2 Elevations Correlate with Persistent Kidney Injury and Progression to Fibrosis
Authors: Arthanarisami, A;Komaru, Y;Katsouridi, C;Schumacher, J;Verges, DK;Ning, L;Abdelmageed, MM;Herrlich, A;Kefaloyianni, E;
Cells
Species: Human
Sample Types: Serum, Urine
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Inflammatory biomarkers at different stages of Sarcopenia in older women
Authors: da Costa Teixeira, LA;Avelar, NCP;Peixoto, MFD;Parentoni, AN;Santos, JMD;Pereira, FSM;Danielewicz, AL;Leopoldino, AAO;Costa, SP;Arrieiro, AN;Soares, LA;da Silva Lage, VK;Prates, ACN;Taiar, R;de Carvalho Bastone, A;Oliveira, VC;Oliveira, MX;Costa, HS;Nobre, JNP;Brant, FP;Duarte, TC;Figueiredo, PHS;Mendonça, VA;Lacerda, ACR;
Scientific reports
Species: Human
Sample Types: Plasma
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Adiponectin Is a Contributing Factor of Low Appendicular Lean Mass in Older Community-Dwelling Women: A Cross-Sectional Study
Authors: LAC Teixeira, JM Dos Santos, AN Parentoni, LP Lima, TC Duarte, FP Brant, CDC Neves, FSM Pereira, NCP Avelar, AL Danielewic, AAO Leopoldino, SP Costa, AN Arrieiro, LA Soares, ACN Prates, JNP Nobre, A de Carvalh, VC de Oliveir, MX Oliveira, PH Scheidt Fi, HS Costa, V Amaral Men, R Taiar, AC Rodrigues
Journal of Clinical Medicine, 2022-12-02;11(23):.
Species: Human
Sample Types: Plasma
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Inflammation-Driven Regulation of PD-L1 and PD-L2, and Their Cross-Interactions with Protective Soluble TNFalpha Receptors in Human Triple-Negative Breast Cancer
Authors: T Baram, N Oren, N Erlichman, T Meshel, A Ben-Baruch
Cancers, 2022-07-19;14(14):.
Species: Human
Sample Types: Cell Culture Supernates
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Adiponectin and Stnfr2 peripheral levels are associated with cardiovascular risk in patients with schizophrenia
Authors: ICS Dias, SM de Campos-, ELM Vieira, APL Mota, PS Azevedo, VTDS Anício, FC Guimarães, LM Mantovani, BF Cruz, AL Teixeira, JV Salgado
Journal of psychiatric research, 2021-11-09;0(0):.
Species: Human
Sample Types: Serum
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Soluble TNF receptors predict acute kidney injury and mortality in critically ill COVID-19 patients: A prospective observational study
Authors: E Sancho Fer, K Hanslin, M Hultström, A Larsson, R Frithiof, M Lipcsey, Uppsala In
Cytokine, 2021-10-05;149(0):155727.
Species: Human
Sample Types: Plasma
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Severe COVID-19 Patients Show an Increase in Soluble TNFR1 and ADAM17, with a Relationship to Mortality
Authors: Y Palacios, A Ruiz, LA Ramón-Luin, R Ocaña-Guzm, O Barreto-Ro, A Sánchez-Mo, B Tecuatzi-C, AG Regalado-G, RD Pineda-Gud, A García-Mar, F Juárez-Her, JP Farias-Con, I Fricke-Gal, G Pérez-Rubi, R Falfán-Val, I Buendia-Ro, K Medina-Que, L Chavez-Gal
International Journal of Molecular Sciences, 2021-08-05;22(16):.
Species: Human
Sample Types: Serum
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Inflammatory and oxidative biomarkers as determinants of functional capacity in patients with COPD assessed by 6-min walk test-derived outcomes
Authors: CDC Neves, VKS Lage, LP Lima, MA Matos, ÉLM Vieira, AL Teixeira, PHS Figueiredo, HS Costa, ACR Lacerda, VA Mendonça
Experimental gerontology, 2021-06-21;152(0):111456.
Species: Human
Sample Types: Plasma
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Exploring the potential effect of paricalcitol on markers of inflammation in de novo renal transplant recipients
Authors: HK Pihlstrøm, T Ueland, AE Michelsen, P Aukrust, F Gatti, C Hammarströ, M Kasprzycka, J Wang, G Haraldsen, G Mjøen, DO Dahle, K Midtvedt, IA Eide, A Hartmann, H Holdaas
PLoS ONE, 2020-12-16;15(12):e0243759.
Species: Human
Sample Types: Plasma
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TNFR1, TNFR2, neutrophil gelatinase-associated lipocalin and heparin binding protein in identifying sepsis and predicting outcome in an intensive care cohort
Authors: M Bergquist, L Samuelsson, A Larsson, J Tydén, J Johansson, M Lipcsey
Scientific Reports, 2020-09-18;10(1):15350.
Species: Human
Sample Types: Plasma
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Predictors of cognitive performance in bipolar disorder: The role of educational degree and inflammatory markers
Authors: IG Barbosa, RA Ferreira, NP Rocha, GC Mol, F da Mata Ch, IE Bauer, AL Teixeira
J Psychiatr Res, 2018-09-17;106(0):31-37.
Species: Human
Sample Types: Plasma
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Effects of the inspiratory muscle training and aerobic training on respiratory and functional parameters, inflammatory biomarkers, redox status and quality of life in hemodialysis patients: A randomized clinical trial
Authors: PHS Figueiredo, MMO Lima, HS Costa, JB Martins, OD Flecha, PF Gonçalves, FL Alves, VGB Rodrigues, EHB Maciel, VA Mendonça, ACR Lacerda, ÉLM Vieira, AL Teixeira, F de Paula, CH Balthazar
PLoS ONE, 2018-07-26;13(7):e0200727.
Species: Human
Sample Types: Plasma
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Periodontal disease influences osteoclastogenic bone markers in subjects with and without rheumatoid arthritis
Authors: J Panezai, A Ghaffar, M Altamash, PE Engström, A Larsson
PLoS ONE, 2018-06-11;13(6):e0197235.
Species: Human
Sample Types: Serum
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TNF receptor signaling is a driver of chronic lymphocytic leukemia that can be therapeutically targeted by the flavonoid wogonin
Authors: C Dürr, BS Hanna, A Schulz, F Lucas, M Zucknick, A Benner, A Clear, S Ohl, S Öztürk, T Zenz, S Stilgenbau, M Li-Weber, PH Krammer, JG Gribben, P Lichter, M Seiffert
Haematologica, 2018-01-11;0(0):.
Species: Human
Sample Types: Serum
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Neutrophil Membrane Cholesterol Content is a Key Factor in Cystic Fibrosis Lung Disease
Authors: MM White, P Geraghty, E Hayes, S Cox, W Leitch, B Alfawaz, GM Lavelle, OJ McElvaney, R Flannery, J Keenan, P Meleady, M Henry, M Clynes, C Gunaratnam, NG McElvaney, EP Reeves
EBioMedicine, 2017-08-16;23(0):173-184.
Species: Human
Sample Types: Plasma
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Neuroendocrine Inflammatory Responses in Overweight/Obese Infants
PLoS ONE, 2016-12-01;11(12):e0167593.
Species: Human
Sample Types: Plasma
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ADAM17 substrate release in proximal tubule drives kidney fibrosis
JCI Insight, 2016-08-18;1(13):.
Species: Human
Sample Types: Urine
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Aqueous two-phase system patterning of detection antibody solutions for cross-reaction-free multiplex ELISA.
Authors: Frampton, John P, White, Joshua B, Simon, Arlyne B, Tsuei, Michael, Paczesny, Sophie, Takayama, Shuichi
Sci Rep, 2014-05-02;4(0):4878.
Species: Human
Sample Types: Plasma
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Bone marrow mesenchymal stromal cells to treat tissue damage in allogeneic stem cell transplant recipients: correlation of biological markers with clinical responses.
Authors: Yin F, Battiwalla M, Ito S, Feng X, Chinian F, Melenhorst J, Koklanaris E, Sabatino M, Stroncek D, Samsel L, Klotz J, Hensel N, Robey P, Barrett A
Stem Cells, 2014-05-01;32(5):1278-88.
Species: Human
Sample Types: Plasma
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Differences in insulin sensitivity, lipid metabolism and inflammation between young adult Pakistani and Norwegian patients with type 2 diabetes: a cross sectional study.
Authors: Wium C, Aasheim E, Ueland T, Michelsen A, Thorsby P, Larsen I, Torjesen P, Aukrust P, Birkeland K
BMC Endocr Disord, 2013-10-22;13(0):49.
Species: Human
Sample Types: Plasma
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Tumor necrosis factor receptor 1 associates with CD137 ligand and mediates its reverse signaling.
Authors: Moh M, Lorenzini P, Gullo C, Schwarz H
FASEB J, 2013-04-25;27(8):2957-66.
Species: Human
Sample Types: Cell Culture Supernates
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A disintegrin and metalloproteinase 17 (ADAM17) and epidermal growth factor receptor (EGFR) signaling drive the epithelial response to Staphylococcus aureus toxic shock syndrome toxin-1 (TSST-1).
Authors: Breshears L, Schlievert P, Peterson M
J Biol Chem, 2012-07-25;287(39):32578-87.
Species: Human
Sample Types: Cell Culture Supernates
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Anti-Tumour Effects of a Specific Anti-ADAM17 Antibody in an Ovarian Cancer Model In Vivo.
Authors: Richards FM, Tape CJ, Jodrell DI, Murphy G
PLoS ONE, 2012-07-11;7(7):e40597.
Species: Human
Sample Types: Tissue Homogenates
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Plasma levels of procalcitonin and eight additional inflammatory molecules in febrile neutropenic patients.
Authors: Neuenschwander LC, Bittencourt H, Ribeiro AF, Teixeira AL, Teixeira MM, Teixeira JC, Nobre V
Clinics (Sao Paulo), 2011-01-01;66(10):1699-705.
Species: Human
Sample Types: Plasma
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Systemic inflammatory markers in COPD: results from the Bergen COPD Cohort Study.
Authors: Eagan TM, Ueland T, Wagner PD, Hardie JA, Mollnes TE, Damas JK, Aukrust P, Bakke PS
Eur. Respir. J., 2009-07-30;35(3):540-8.
Species: Human
Sample Types: Plasma
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Effects of ciliary muscle plasmid electrotransfer of TNF-alpha soluble receptor variants in experimental uveitis.
Authors: Touchard E, Bloquel C, Bigey P, Kowalczuc L, Jonet L, Thillaye-Goldenberg B, Naud MC, Scherman D, de Kozak Y, Benezra D, Behar-Cohen F
Gene Ther., 2009-05-14;16(7):862-73.
Species: Human
Sample Types: Recombinant Protein
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A tumor necrosis factor-alpha-mediated pathway promoting autosomal dominant polycystic kidney disease.
Authors: Li X, Magenheimer BS, Xia S, Johnson T, Wallace DP, Calvet JP, Li R
Nat. Med., 2008-06-15;14(8):863-8.
Species: Human
Sample Types: Urine
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Suspension microarrays for the identification of the response patterns in hyperinflammatory diseases.
Authors: Hsu HY, Wittemann S, Schneider EM, Weiss M, Joos TO
Med Eng Phys, 2008-03-03;30(8):976-83.
Species: Human
Sample Types: Plasma
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Cutting edge: TNFR-shedding by CD4+CD25+ regulatory T cells inhibits the induction of inflammatory mediators.
Authors: van Mierlo GJ, Scherer HU, Hameetman M, Morgan ME, Flierman R, Huizinga TW, Toes RE
J. Immunol., 2008-03-01;180(5):2747-51.
Species: Human
Sample Types: Cell Culture Supernates
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Circulating levels of tumor necrosis factor receptors are highly predictive of mortality in patients with rheumatoid arthritis.
Authors: Mattey DL, Glossop JR, Nixon NB, Dawes PT
Arthritis Rheum., 2007-12-01;56(12):3940-8.
Species: Human
Sample Types: Serum
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Elevated urinary VCAM-1, P-selectin, soluble TNF receptor-1, and CXC chemokine ligand 16 in multiple murine lupus strains and human lupus nephritis.
Authors: Wu T, Xie C, Wang HW, Zhou XJ, Schwartz N, Calixto S, Mackay M, Aranow C, Putterman C, Mohan C
J. Immunol., 2007-11-15;179(10):7166-75.
Species: Human
Sample Types: Urine
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Early cytokine responses after percutaneous magnetic resonance imaging guided laser thermoablation of malignant liver tumors.
Authors: Kallio R, Sequeiros R, Surcel HM, Ohtonen P, Kiviniemi H, Syrjala H
Cytokine, 2006-07-25;34(5):278-83.
Species: Human
Sample Types: Serum
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Polymorphism in the tumour necrosis factor receptor II gene is associated with circulating levels of soluble tumour necrosis factor receptors in rheumatoid arthritis.
Authors: Glossop JR, Dawes PT, Nixon NB, Mattey DL
Arthritis Res. Ther., 2005-09-07;7(6):R1227-34.
Species: Human
Sample Types: Serum
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