Human TNF RI/TNFRSF1A DuoSet ELISA

Catalog # Availability Size / Price Qty
DY225
DY225-05
Ancillary Products Available
Human TNF RI / TNFRSF1A ELISA Standard Curve
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Product Details
Procedure
Citations (23)
FAQs
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Human TNF RI/TNFRSF1A DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For five or 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 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)

 

  

Data Example

Human TNF RI / TNFRSF1A ELISA Standard Curve

Product Datasheets

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: 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.

Long Name:
Tumor Necrosis Factor Receptor I
Entrez Gene IDs:
7132 (Human); 21937 (Mouse); 25625 (Rat); 102135285 (Cynomolgus Monkey)
Alternate Names:
CD120a antigen; CD120a; FPF; p55; p55-R; p60; TNF RI; TNFARMGC19588; TNF-R; TNF-R1; TNFR1TBP1; TNFR55; TNF-R55; TNFR60; TNFRI; TNF-RI; TNF-R-I; TNFR-I; TNFRSF1A; tumor necrosis factor binding protein 1; Tumor necrosis factor receptor 1; tumor necrosis factor receptor 1A isoform beta; tumor necrosis factor receptor superfamily member 1A; tumor necrosis factor receptor superfamily, member 1A; tumor necrosis factor receptor type 1; Tumor necrosis factor receptor type I; tumor necrosis factor-alpha receptor

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, 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 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.

23 Citations: Showing 1 - 10
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  1. 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;106(0):31-37.
    Species: Human
    Sample Types: Plasma
  2. 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;13(7):e0200727.
    Species: Human
    Sample Types: Plasma
  3. 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;13(6):e0197235.
    Species: Human
    Sample Types: Serum
  4. 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;0(0):.
    Species: Human
    Sample Types: Serum
  5. 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;23(0):173-184.
    Species: Human
    Sample Types: Plasma
  6. ADAM17 substrate release in proximal tubule drives kidney fibrosis
    JCI Insight, 2016;1(13):.
    Species: Human
    Sample Types: Urine
  7. Neuroendocrine Inflammatory Responses in Overweight/Obese Infants
    PLoS ONE, 2016;11(12):e0167593.
    Species: Human
    Sample Types: Plasma
  8. 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;4(0):4878.
    Species: Human
    Sample Types: Plasma
  9. 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;32(5):1278-88.
    Species: Human
    Sample Types: Plasma
  10. 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;27(8):2957-66.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. 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;13(0):49.
    Species: Human
    Sample Types: Plasma
  12. 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;287(39):32578-87.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. 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;7(7):e40597.
    Species: Human
    Sample Types: Tissue Homogenates
  14. 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;66(10):1699-705.
    Species: Human
    Sample Types: Plasma
  15. 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., 2010;35(3):540-8.
    Species: Human
    Sample Types: Plasma
  16. 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;16(7):862-73.
    Species: Human
    Sample Types: Recombinant Protein
  17. 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;14(8):863-8.
    Species: Human
    Sample Types: Urine
  18. 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;30(8):976-83.
    Species: Human
    Sample Types: Plasma
  19. 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;180(5):2747-51.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. 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;179(10):7166-75.
    Species: Human
    Sample Types: Urine
  21. 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, 2007;34(5):278-83.
    Species: Human
    Sample Types: Serum
  22. 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;56(12):3940-8.
    Species: Human
    Sample Types: Serum
  23. 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;7(6):R1227-34.
    Species: Human
    Sample Types: Serum

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