hh Human sTNF RII/TNFRSF1B Quantikine ELISA Kit DRT200: R&D Systems

Human sTNF RII/TNFRSF1B Quantikine ELISA Kit

(48 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    3.5 hours or 4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (200 uL), Serum (20 uL), EDTA Plasma (20 uL), Heparin Plasma (20 uL), Citrate Plasma (20 uL), Urine (20 uL)
  • Sensitivity
    2.3 pg/mL
  • Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
  • Specificity
    Natural and recombinant human sTNF RII
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC05, Quantikine Immunoassay Control Group 2 - Please Inquire
Product Summary
The Quantikine Human sTNF RII Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure human sTNF RII in cell culture supernates, serum, plasma, or urine. It contains E. coli-expressed recombinant human sTNF RII as well as antibodies raised against this polypeptide. The recombinant protein represents the non-glycosylated, N-terminal methionyl form of the naturally occurring human soluble Type II receptor for TNF, minus 53 amino acids from the proline-rich region of the protein just exterior to the transmembrane domain, with an apparent molecular weight of approximately 19.8 kDa. This immunoassay has been shown to accurately quantitate the recombinant soluble sTNF RII. Results obtained using natural sTNF RII showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural sTNF RII. Since the measurement of human sTNF RII by this immunoassay is insensitive to added TNF-alpha or TNF-beta, it is probable that this measurement corresponds to the total amount of the soluble receptor present in samples, (i.e. the total amount of free receptor plus the total amount of receptor bound to TNF).

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean68.717934989.8197444
Standard Deviation2.24.616.84.66.916.2
CV%3.22.64.85.13.53.6

Cell Culture Supernates, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean45.312625497.1199430
Standard Deviation23.710.43.16.917.8
CV%4.42.94.13.23.54.1

Recovery

The recovery of human sTNF RII spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 96 88-103
Citrate Plasma (n=4) 100 91-116
EDTA Plasma (n=4) 95 85-100
Heparin Plasma (n=4) 89 74-108
Serum (n=4) 97 86-107
Urine (n=4) 97 83-111
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of sTNF RII were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human sTNF RII/TNFRSF1B Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
    • Long Name
      Tumor Necrosis Factor Receptor II
    • Entrez Gene IDs
      7133 (Human); 21938 (Mouse); 156767 (Rat);
    • Alternate Names
      CD120b antigen; CD120b; Etanercept; p75 TNF receptor; p75TBPII; p75TNFR; soluble TNFR1B variant 1; TNF RII; TNFBRp80 TNF-alpha receptor; TNF-R2; TNFR2TNFR1B; TNF-R75; TNFR80; TNFRII; TNF-RII; TNF-R-II; TNFR-II; TNFRSF1B; tumor necrosis factor beta receptor; tumor necrosis factor binding protein 2; Tumor necrosis factor receptor 2; tumor necrosis factor receptor superfamily member 1B; tumor necrosis factor receptor superfamily, member 1B; Tumor necrosis factor receptor type II;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 50 µL Assay Diluent
    4.   Add 50 µL of Assay Diluent to each well.

    5. 200 µL Standard, Control, or Sample
    6.   Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    7.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well.
    10.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
      For Cell Culture Supernate Samples & Urine Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
    11.   Aspirate and wash 3 times.

    12. 200 µL Substrate Solution
    13.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.

    14. 50 µL Stop Solution
    15. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    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.

    48 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Circulating TNF Receptors 1 and 2 Predict Mortality in Patients with End-stage Renal Disease Undergoing Dialysis
      Authors: T Gohda, S Maruyama, N Kamei, S Yamaguchi, T Shibata, M Murakoshi, S Horikoshi, Y Tomino, I Ohsawa, H Gotoh, S Nojiri, Y Suzuki
      Sci Rep, 2017;7(0):43520.
      Species: Human
      Sample Type: Serum
    2. Anti-TNF-? Drugs Differently Affect the TNF?-sTNFR System and Monocyte Subsets in Patients with Psoriasis
      PLoS ONE, 2016;11(12):e0167757.
      Species: Human
      Sample Type: Plasma
    3. Altered Monocyte and Endothelial Cell Adhesion Molecule Expression Is Linked to Vascular Inflammation in Human Immunodeficiency Virus Infection
      Authors: M Kulkarni, E Bowman, J Gabriel, T Amburgy, E Mayne, DA Zidar, C Maierhofer, AN Turner, JA Bazan, SL Koletar, MM Lederman, SF Sieg, NT Funderburg
      Open Forum Infect Dis, 2016;3(4):ofw224.
      Species: Human
      Sample Type: Plasma
    4. Local TNF causes NFATc1-dependent cholesterol-mediated podocyte injury
      J Clin Invest, 2016;0(0):.
      Species: Human
      Sample Type: Serum
    5. Inflammatory Markers and the Risk of Hip and Vertebral Fractures in Men: the Osteoporotic Fractures in Men (MrOS)
      J Bone Miner Res, 2016;0(0):.
      Species: Human
      Sample Type: Serum
    6. Stability of cytokines, chemokines and soluble activation markers in unprocessed blood stored under different conditions
      Cytokine, 2016;84(0):17-24.
      Species: Human
      Sample Type: Plasma
    7. Association between serum soluble tumor necrosis factor-? receptors and early childhood obesity
      Authors: S Ouyang, W Li, Z Liu, Y Li, S Li, J Wu
      Endocr J, 2016;0(0):.
      Species: Human
      Sample Type: Serum
    8. Anti-Drug Antibodies, Drug Levels, Interleukin-6 and Soluble TNF Receptors in Rheumatoid Arthritis Patients during the First 6 Months of Treatment with Adalimumab or Infliximab: A Descriptive Cohort Study
      PLoS ONE, 2016;11(9):e0162316.
      Species: Human
      Sample Type: Serum
    9. Reliability of Potential Pain Biomarkers in the Saliva of Healthy Subjects: Inter-Individual Differences and Intersession Variability
      PLoS ONE, 2016;11(12):e0166976.
      Species: Human
      Sample Type: Saliva
    10. Prevalent peripheral arterial disease and inflammatory burden
      BMC Geriatr, 2016;16(1):213.
      Species: Human
      Sample Type: Serum
    11. Late Antiretroviral Therapy (ART) Initiation Is Associated with Long-Term Persistence of Systemic Inflammation and Metabolic Abnormalities.
      Authors: Ghislain M, Bastard J, Meyer L, Capeau J, Fellahi S, Gerard L, May T, Simon A, Vigouroux C, Goujard C
      PLoS ONE, 2015;10(12):e0144317.
      Species: Human
      Sample Type: Serum
    12. Plasma TNF-alpha and Soluble TNF Receptor Levels after Doxorubicin with or without Co-Administration of Mesna-A Randomized, Cross-Over Clinical Study.
      Authors: Hayslip J, Dressler E, Weiss H, Taylor T, Chambers M, Noel T, Miriyala S, Keeney J, Ren X, Sultana R, Vore M, Butterfield D, St Clair D, Moscow J
      PLoS ONE, 2015;10(4):e0124988.
      Species: Human
      Sample Type: Plasma
    13. Circulating TNF receptors 1 and 2 are associated with the severity of renal interstitial fibrosis in IgA nephropathy.
      Authors: Sonoda Y, Gohda T, Suzuki Y, Omote K, Ishizaka M, Matsuoka J, Tomino Y
      PLoS ONE, 2015;10(4):e0122212.
      Species: Human
      Sample Type: Serum
    14. Circulating levels of tumor necrosis factor-alpha receptor 2 are increased in heart failure with preserved ejection fraction relative to heart failure with reduced ejection fraction: evidence for a divergence in pathophysiology.
      Authors: Putko B, Wang Z, Lo J, Anderson T, Becher H, Dyck J, Kassiri Z, Oudit G
      PLoS ONE, 2014;9(6):e99495.
      Species: Human
      Sample Type: Plasma
    15. A case-control study of pre-operative levels of serum neutrophil gelatinase-associated lipocalin and other potential inflammatory markers in colorectal cancer.
      Authors: Duvillard L, Ortega-Deballon P, Bourredjem A, Scherrer M, Mantion G, Delhorme J, Deguelte-Lardiere S, Petit J, Bonithon-Kopp C
      BMC Cancer, 2014;14(0):912.
      Species: Human
      Sample Type: Serum
    16. Synergism between circulating tumor necrosis factor receptor 2 and HbA(1c) in determining renal decline during 5-18 years of follow-up in patients with type 1 diabetes and proteinuria.
      Authors: Skupien J, Warram J, Niewczas M, Gohda T, Malecki M, Mychaleckyj J, Galecki A, Krolewski A
      Diabetes Care, 2014;37(9):2601-8.
      Species: Human
      Sample Type: Serum
    17. Effect of IL-6 on the insulin sensitivity in patients with type 2 diabetes.
      Authors: Harder-Lauridsen N, Krogh-Madsen R, Holst J, Plomgaard P, Leick L, Pedersen B, Fischer C
      Am J Physiol Endocrinol Metab, 2014;306(7):E769-78.
      Species: Human
      Sample Type: Plasma
    18. Negative and competitive social interactions are related to heightened proinflammatory cytokine activity.
      Authors: Chiang JJ, Eisenberger NI, Seeman TE, Taylor SE
      Proc. Natl. Acad. Sci. U.S.A., 2012;109(6):1878-82.
      Species: Human
      Sample Type: Plasma
    19. Compensated inflammation in systemic juvenile idiopathic arthritis: role of alternatively activated macrophages.
      Authors: Shimizu M, Yachie A
      Cytokine, 2012;60(1):226-32.
      Species: Human
      Sample Type: Serum
    20. Inflammation marker and risk of pancreatic cancer: a nested case-control study within the EPIC cohort.
      Authors: Grote VA, Kaaks R, Nieters A, Tjonneland A, Halkjaer J, Overvad K, Skjelbo Nielsen MR, Boutron-Ruault MC, Clavel-Chapelon F, Racine A, Teucher B, Becker S, Pischon T, Boeing H, Trichopoulou A, Cassapa C, Stratigakou V, Palli D, Krogh V, Tumino R, Vineis P, Panico S, Rodriguez L, Duell EJ, Sanchez MJ, Dorronsoro M, Navarro C, Gurrea AB, Siersema PD, Peeters PH, Ye W, Sund M, Lindkvist B, Johansen D, Khaw KT, Wareham N, Allen NE, Travis RC, Fedirko V, Jenab M, Michaud DS, Chuang SC, Romaguera D, Bueno-de-Mesquita HB, Rohrmann S
      Br. J. Cancer, 2012;106(11):1866-74.
      Species: Human
      Sample Type: Serum
    21. Inflammatory markers and the risk of hip fracture: the Women's Health Initiative.
      Authors: Barbour KE, Boudreau R, Danielson ME, Youk AO, Wactawski-Wende J, Greep NC, LaCroix AZ, Jackson RD, Wallace RB, Bauer DC, Allison MA, Cauley JA
      J. Bone Miner. Res., 2012;27(5):1167-76.
      Species: Human
      Sample Type: Serum
    22. Elevated CSF levels of TACE activity and soluble TNF receptors in subjects with mild cognitive impairment and patients with Alzheimer's disease.
      Authors: Jiang H, Hampel H, Prvulovic D, Wallin A, Blennow K, Li R, Shen Y
      Mol Neurodegener, 2011;6(0):69.
      Species: Human
      Sample Type: CSF
    23. Procalcitonin as an early marker of bacterial infection in neutropenic febrile children with acute lymphoblastic leukemia.
      Authors: Hatzistilianou M, Rekliti A, Athanassiadou F, Catriu D
      Inflamm. Res., 2010;59(5):339-47.
      Species: Human
      Sample Type: Serum
    24. Neurobiological correlates of coping through emotional approach.
      Authors: Master SL, Amodio DM, Stanton AL, Yee CM, Hilmert CJ, Taylor SE
      Brain Behav. Immun., 2009;23(1):27-35.
      Species: Human
      Sample Type: Saliva
    25. Cross-sectional relations of multiple inflammatory biomarkers to peripheral arterial disease: The Framingham Offspring Study.
      Authors: Murabito JM, Keyes MJ, Guo CY, Keaney JF, Vasan RS, D'Agostino RB, Benjamin EJ
      Atherosclerosis, 2009;203(2):509-14.
      Species: Human
      Sample Type: Plasma
    26. Biomarkers of inflammation and development of rheumatoid arthritis in women from two prospective cohort studies.
      Authors: Karlson EW, Chibnik LB, Tworoger SS, Lee IM, Buring JE, Shadick NA, Manson JE, Costenbader KH
      Arthritis Rheum., 2009;60(3):641-52.
      Species: Human
      Sample Type: Plasma
    27. Adrenomedullin and endothelin-1 are related to inflammation in chronic heart failure.
      Authors: Gombos T, Forhecz Z, Pozsonyi Z, Wallentin S, Papassotiriou J, Kunde J, Morgenthaler NG, Janoskuti L, Prohaszka Z
      Inflamm. Res., 2009;58(6):298-305.
      Species: Human
      Sample Type: Serum
    28. Comparing Type D personality and older age as correlates of tumor necrosis factor-alpha dysregulation in chronic heart failure.
      Authors: Denollet J, Vrints CJ, Conraads VM
      Brain Behav. Immun., 2008;22(5):736-43.
      Species: Human
      Sample Type: Plasma
    29. Serum interleukin-18 and soluble tumour necrosis factor receptor 2 are associated with disease severity in patients with paracoccidioidomycosis.
      Authors: Corvino CL, Mamoni RL, Fagundes GZ, Blotta MH
      Clin. Exp. Immunol., 2007;147(3):483-90.
      Species: Human
      Sample Type: Serum
    30. Muscle atrophy and hypertrophy signaling in patients with chronic obstructive pulmonary disease.
      Authors: Doucet M, Russell AP, Leger B, Debigare R, Joanisse DR, Caron MA, LeBlanc P, Maltais F
      Am. J. Respir. Crit. Care Med., 2007;176(3):261-9.
      Species: Human
      Sample Type: Plasma
    31. Selective functional exhaustion of hematopoietic progenitor cells in the bone marrow of patients with postinfarction heart failure.
      Authors: Kissel CK, Lehmann R, Assmus B, Aicher A, Honold J, Fischer-Rasokat U, Heeschen C, Spyridopoulos I, Dimmeler S, Zeiher AM
      J. Am. Coll. Cardiol., 2007;49(24):2341-9.
      Species: Human
      Sample Type: Serum
    32. Inflammatory markers and incident fracture risk in older men and women: the Health Aging and Body Composition Study.
      Authors: Cauley JA, Danielson ME, Boudreau RM, Forrest KY, Zmuda JM, Pahor M, Tylavsky FA, Cummings SR, Harris TB, Newman AB
      J. Bone Miner. Res., 2007;22(7):1088-95.
      Species: Human
      Sample Type: Serum
    33. The shedding activity of ADAM17 is sequestered in lipid rafts.
      Authors: Tellier E, Rebsomen L
      Exp. Cell Res., 2006;312(20):3969-80.
      Species: Primate - Cholrocebus pygerythrus (Vervet Monkey)
      Sample Type: Cell Lysates
    34. Human intestinal intraepithelial lymphocytes keep TNF alpha levels low by cell uptake and feedback inhibition of transcription.
      Authors: Ebert EC, Mehta V
      Cell. Immunol., 2006;241(1):7-13.
      Species: Human
      Sample Type: Cell Culture Supernates
    35. Proinflammatory cytokines, insulin resistance, and insulin secretion in chronic hepatitis C patients: A case-control study.
      Authors: Lecube A, Hernandez C, Genesca J, Simo R
      Diabetes Care, 2006;29(5):1096-101.
      Species: Human
      Sample Type: Serum
    36. Analysis of tumor necrosis factor-alpha, interleukin-6, interleukin-1beta, soluble tumor necrosis factor receptors I and II, interleukin-6 soluble receptor, interleukin-1 soluble receptor type II, interleukin-1 receptor antagonist, and protein in the synovial fluid of patients with temporomandibular joint disorders.
      Authors: Kaneyama K, Segami N, Sun W, Sato J, Fujimura K
      Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2005;99(3):276-84.
      Species: Human
      Sample Type: Synovial Fluid
    37. Growth hormone-induced reduction of soluble apoptosis mediators is associated with reverse cardiac remodelling and improvement of exercise capacity in patients with idiopathic dilated cardiomyopathy.
      Authors: Parissis JT, Adamopoulos S, Karatzas D, Paraskevaidis J, Livanis E, Kremastinos D
      Eur J Cardiovasc Prev Rehabil, 2005;12(2):164-8.
      Species: Human
      Sample Type: Plasma
    38. Changes in HIV RNA viral load, CD4+ T-cell counts, and levels of immune activation markers associated with anti-tuberculosis therapy and cotrimoxazole prophylaxis among HIV-infected tuberculosis patients in Abidjan, Cote d'Ivoire.
      Authors: Kalou M, Sassan-Morokro M, Abouya L, Bile C, Maurice C, Maran M, Tossou O, Roels T, Greenberg AE, Wiktor SZ, Nkengasong JN
      J. Med. Virol., 2005;75(2):202-8.
      Species: Human
      Sample Type: Serum
    39. Levels of soluble cytokine factors in temporomandibular joint effusions seen on magnetic resonance images.
      Authors: Kaneyama K, Segami N, Sun W, Sato J, Fujimura K
      Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2005;99(4):411-8.
      Species: Human
      Sample Type: Synovial Fluid
    40. Experimental heatstroke in baboon: analysis of the systemic inflammatory response.
      Authors: Bouchama A, Ollivier V, Roberts G, Al Mohanna F, de Prost D, Eldali A, Saussereau E, El-Sayed R, Chollet-Martin S
      Shock, 2005;24(4):332-5.
      Species: Primate - Papio hamadryas (Hamadyras Baboon)
      Sample Type: Plasma
    41. Regulation of peritoneal and systemic neutrophil-derived tumor necrosis factor-alpha release in patients with severe peritonitis: role of tumor necrosis factor-alpha converting enzyme cleavage.
      Authors: Kermarrec N, Selloum S, Plantefeve G, Chosidow D, Paoletti X, Lopez A, Mantz J, Desmonts JM, Gougerot-Pocidalo MA, Chollet-Martin S
      Crit. Care Med., 2005;33(6):1359-64.
      Species: Human
      Sample Type: Cell Culture Supernates
    42. Pro- and anti-inflammatory responses in respiratory syncytial virus bronchiolitis.
      Authors: McNamara PS, Flanagan BF, Selby AM, Hart CA, Smyth RL
      Eur. Respir. J., 2004;23(1):106-12.
      Species: Human
      Sample Type: BALF
    43. Immunological effects of induced shame and guilt.
      Authors: Dickerson SS, Kemeny ME, Aziz N, Kim KH, Fahey JL
      Psychosom Med, 2004;66(1):124-31.
      Species: Human
      Sample Type: Saliva
    44. Arenavirus-mediated liver pathology: acute lymphocytic choriomeningitis virus infection of rhesus macaques is characterized by high-level interleukin-6 expression and hepatocyte proliferation.
      Authors: Lukashevich IS, Tikhonov I, Rodas JD, Zapata JC, Yang Y, Djavani M, Salvato MS
      J. Virol., 2003;77(3):1727-37.
      Species: Primate - Macaca mulatta (Rhesus Macaque)
      Sample Type: Serum
    45. Functional and phenotypic changes in circulating lymphocytes from hospitalized zambian children with measles.
      Authors: Ryon JJ, Moss WJ, Monze M, Griffin DE
      Clin. Diagn. Lab. Immunol., 2002;9(5):994-1003.
      Species: Human
      Sample Type: Plasma
    46. Age-related autoantibody production in a nonhuman primate model.
      Authors: Attanasio R, Brasky KM, Robbins SH, Jayashankar L, Nash RJ, Butler TM
      Clin. Exp. Immunol., 2001;123(3):361-5.
      Species: Primate - Papio cynocephalus (Yellow Baboon)
      Sample Type: Serum
    47. Tumour necrosis factor alpha and its soluble receptors in juvenile chronic arthritis.
      Authors: Rooney M, Varsani H, Martin K, Lombard PR, Dayer JM, Woo P
      Rheumatology (Oxford), 2000;39(4):432-8.
      Species: Human
      Sample Type: Serum
    48. An analysis of 14 molecular markers for monitoring osteoarthritis: segregation of the markers into clusters and distinguishing osteoarthritis at baseline.
      Authors: Otterness IG, Swindell AC, Zimmerer RO, Poole AR, Ionescu M, Weiner E
      Osteoarthr. Cartil., 2000;8(3):180-5.
      Species: Human
      Sample Type: Serum
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