Human sTNF RII/TNFRSF1B 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 TNF RII/TNFRSF1B. 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: TNF RII/TNFRSF1B
TNF RII (Tumor Necrosis Factor Receptor II), also known as TNFRSF1B, p75/p80, and CD120b, is a widely expressed receptor for membrane-associated TNF-alpha and Lymphotoxin-alpha. Its activation initiates pro-inflammatory and pro-survival responses via NFkB-dependent signaling pathways, although it may also induce apoptosis.
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 sTNF RII/TNFRSF1B 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.
27
Citations: Showing 1 - 10
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Advancing Point-of-Care Diagnosis: Digitalizing Combinatorial Biomarker Signals for Lupus Nephritis
Authors: Guo, J;Teymur, A;Tang, C;Saxena, R;Wu, T;
Biosensors
Species: Human
Sample Types: Serum
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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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Antibody microarray analysis of amniotic fluid proteomes in women with cervical insufficiency and short cervix, and their association with pregnancy latency length
Authors: S Hong, KH Park, YE Lee, JE Lee, YM Kim, E Joo, I Cho
PLoS ONE, 2022-02-07;17(2):e0263586.
Species: Human
Sample Types: Amniotic Fluid
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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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Neurocognitive outcomes in Malawian children exposed to malaria during pregnancy: An observational birth cohort study
Authors: AM Weckman, AL Conroy, M Madanitsa, B Gnaneswara, CR McDonald, L Kalilani-P, J Chandna, D Ali, V Mwapasa, C Khairallah, KL Thwai, SR Meshnick, SM Taylor, FO Ter Kuile, KC Kain, M Gladstone
PLoS medicine, 2021-09-28;18(9):e1003701.
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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Persistence of monocyte activation under treatment in people followed since acute HIV-1 infection relative to participants at high or low risk of HIV infection
Authors: S Novelli, C Lécuroux, C Goujard, J Reynes, A Villemant, L Blum, A Essat, V Avettand-F, O Launay, JM Molina, C Bourgeois, L Meyer
EBioMedicine, 2020-11-26;62(0):103129.
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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Systemic inflammation is associated with malaria and preterm birth in women living with HIV on antiretrovirals and co-trimoxazole
Authors: CR McDonald, AM Weckman, AL Conroy, P Olwoch, P Natureeba, MR Kamya, DV Havlir, G Dorsey, KC Kain
Sci Rep, 2019-05-01;9(1):6758.
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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Neuroendocrine Inflammatory Responses in Overweight/Obese Infants
PLoS ONE, 2016-12-01;11(12):e0167593.
Species: Human
Sample Types: Plasma
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Axl, Ferritin, Insulin-Like Growth Factor Binding Protein 2, and Tumor Necrosis Factor Receptor Type II as Biomarkers in Systemic Lupus Erythematosus
Authors: CC Mok, HH Ding, M Kharboutli, C Mohan
Arthritis Care Res (Hoboken), 2016-07-28;68(9):1303-9.
Species: Human
Sample Types: Serum
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Inflammatory and Angiogenic Factors at Mid-Pregnancy Are Associated with Spontaneous Preterm Birth in a Cohort of Tanzanian Women.
Authors: McDonald C, Darling A, Conroy A, Tran V, Cabrera A, Liles W, Wang M, Aboud S, Urassa W, Fawzi W, Kain K
PLoS ONE, 2015-08-06;10(8):e0134619.
Species: Human
Sample Types: Plasma
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Soluble TNF receptors and kidney dysfunction in the elderly.
Authors: Carlsson A, Larsson T, Helmersson-Karlqvist J, Larsson A, Lind L, Arnlov J
J Am Soc Nephrol, 2014-02-07;25(6):1313-20.
Species: Human
Sample Types: Whole Blood
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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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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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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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Immunomodulating effects of CKBM on the cytokine production in peripheral blood mononuclear cells (PBMCs) from healthy volunteers.
Authors: Chan BP, Yuen WF, Lee WH, Wong SN, Chung TY, Wu YJ, Pang SF
Immunopharmacol Immunotoxicol, 2004-05-01;26(2):177-92.
Species: Human
Sample Types: Cell Culture Supernates
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Adherence influences monocyte responsiveness to interleukin-10.
Authors: Petit-Bertron AF, Fitting C, Cavaillon JM, Adib-Conquy M
J. Leukoc. Biol., 2003-01-01;73(1):145-54.
Species: Human
Sample Types: Cell Culture Supernates
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