Human CD25/IL-2R alpha Quantikine ELISA Kit

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CD25/IL-2R alpha ELISA Kit ELISA
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Human CD25/IL-2R alpha Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (10 uL), Serum (13 uL), EDTA Plasma (13 uL), Heparin Plasma (13 uL)
Sensitivity
10 pg/mL
Assay Range
78.0 - 5,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human IL-2 sR alpha
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human IL-2 sR alpha Immunoassay is a 3.5 hour solid phase ELISA designed to measure IL-2 sR alpha in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human IL-2 sR alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IL-2 sR alpha showed linear curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-2 sR alpha.

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

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 207 613 2357 188 630 2615
Standard Deviation 12.7 37.5 108 13.6 38.1 183
CV% 6.1 6.1 4.6 7.2 6 7

Recovery

The recovery of IL-2 sR alpha spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 105 93-118
EDTA Plasma (n=5) 103 91-113
Heparin Plasma (n=5) 104 98-113
Serum (n=5) 105 99-116

Linearity

To assess the linearity of the assay, five samples containing or spiked with high concentrations of IL-2 sR alpha in various matrices were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CD25/IL-2 R alpha ELISA Linearity

Data Examples

Human CD25/IL-2 R alpha ELISA Standard Curve

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CD25/IL-2R alpha

CD25/IL-2 receptor alpha is a transmembrane protein that associates with IL-2 R beta/CD122 and the Common gamma Chain/CD132 on activated T cells and regulatory T cells (Treg). The Common gamma chain is a shared subunit in the receptors for IL-4, -7, -9, -15, and -21. IL-2 R alpha is required for mediating IL-2 induced effects including T cell proliferation, activation induced cell death (AICD) of autoreactive naive T cells, and the development of CD4+CD25+ Treg which contribute to peripheral T cell homeostasis. A soluble form of IL-2 R alpha can be generated by proteolytic cleavage of the cell surface receptor.

Long Name:
Interleukin 2 Receptor alpha
Entrez Gene IDs:
3559 (Human); 16184 (Mouse); 25704 (Rat); 403870 (Canine)
Alternate Names:
CD25 antigen; CD25; IDDM10; IL-2 R alpha; IL-2 receptor subunit alpha; IL2R alpha; IL-2R subunit alpha; IL2R; IL2RA; IL-2Ra; IL-2-RA; IL2-RA; interleukin 2 receptor, alpha; interleukin-2 receptor subunit alpha; p55; TAC antigen; TCGFR
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Proceed to the next step. Do not incubate or wash.

  7. 100 µL Conjugate
  8.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 3 hours.
  9.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  10. 200 µL Substrate Solution
  11.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.

  12. 50 µL Stop Solution
  13.   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 for Human CD25/IL-2R alpha Quantikine ELISA Kit

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.

29 Citations: Showing 1 - 10
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  1. Germline HAVCR2 mutations altering TIM-3 characterize subcutaneous panniculitis-like T cell lymphomas with hemophagocytic lymphohistiocytic syndrome
    Authors: T Gayden, FE Sepulveda, DA Khuong-Qua, J Pratt, ET Valera, A Garrigue, S Kelso, F Sicheri, LG Mikael, N Hamel, A Bajic, R Dali, S Deshmukh, D Dervovic, D Schramek, F Guerin, M Taipale, H Nikbakht, J Majewski, D Moshous, J Charlebois, S Abish, C Bole-Feyso, P Nitschke, B Bader-Meun, D Mitchell, C Thieblemon, M Battistell, S Gravel, VH Nguyen, R Conyers, JS Diana, C McCormack, HM Prince, M Besnard, S Blanche, PG Ekert, S Fraitag, WD Foulkes, A Fischer, B Neven, D Michonneau, G de Saint B, N Jabado
    Nat. Genet., 2018;0(0):.
    Species: Human
    Sample Types: Serum
  2. Reevaluation of immune activation in the era of cART and an aging HIV-infected population
    Authors: LR de Armas, S Pallikkuth, V George, S Rinaldi, R Pahwa, KL Arheart, S Pahwa
    JCI Insight, 2017;2(20):.
    Species: Human
    Sample Types: Plasma
  3. Incidence and Risk Factors for Developing Dengue-Associated Hemophagocytic Lymphohistiocytosis in Puerto Rico, 2008 - 2013
    PLoS Negl Trop Dis, 2016;10(8):e0004939.
    Species: Human
    Sample Types: Serum
  4. Stability of cytokines, chemokines and soluble activation markers in unprocessed blood stored under different conditions
    Cytokine, 2016;84(0):17-24.
    Species: Human
    Sample Types: Plasma
  5. Efficacy, durability, and response predictors of low-dose interleukin-2 therapy for chronic graft vs. host disease
    Authors: J Koreth, HT Kim, KT Jones, PB Lange, CG Reynolds, MJ Chammas, K Dusenbury, J Whangbo, S Nikiforow, EP Alyea, P Armand, CS Cutler, VT Ho, YB Chen, D Avigan, BR Blazar, JH Antin, J Ritz, RJ Soiffer
    Blood, 2016;0(0):.
    Species: Human
    Sample Types: Plasma
  6. Augmented efficacy with the combination of blockade of the Notch-1 pathway, bortezomib and romidepsin in a murine MT-1 adult T-cell leukemia model.
    Authors: Yu P, Petrus M, Ju W, Zhang M, Conlon K, Nakagawa M, Maeda M, Bamford R, Waldmann T
    Leukemia, 2015;29(3):556-66.
    Species: Human
    Sample Types: Serum
  7. Elevated levels of CXCL10 in the Periodic Fever, Aphthous stomatitis, Pharyngitis and cervical Adenitis syndrome (PFAPA) during and between febrile episodes; an indication of a persistent activation of the innate immune system.
    Authors: Forsvoll J, Kristoffersen E, Oymar K
    Pediatr Rheumatol Online J, 2013;11(1):38.
    Species: Human
    Sample Types: Plasma
  8. Malignant transformation of CD4+ T lymphocytes mediated by oncogenic kinase NPM/ALK recapitulates IL-2-induced cell signaling and gene expression reprogramming.
    Authors: Marzec M, Halasa K, Liu X, Wang H, Cheng M, Baldwin D, Tobias J, Schuster S, Woetmann A, Zhang Q, Turner S, Odum N, Wasik M
    J Immunol, 2013;191(12):6200-7.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Human CD1c (BDCA-1)+ myeloid dendritic cells secrete IL-10 and display an immuno-regulatory phenotype and function in response to Escherichia coli.
    Authors: Kassianos AJ, Hardy MY, Ju X, Vijayan D, Ding Y, Vulink AJ, McDonald KJ, Jongbloed SL, Wadley RB, Wells C, Hart DN, Radford KJ
    Eur. J. Immunol., 2012;42(6):1512-22.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Synergistic antitumor activity of anti-CD25 recombinant immunotoxin LMB-2 with chemotherapy.
    Authors: Singh R, Zhang Y, Pastan I, Kreitman RJ
    Clin. Cancer Res., 2012;18(1):152-60.
    Species: Human
    Sample Types: Serum
  11. A novel role for IL-22R1 as a driver of inflammation.
    Authors: Savan R, McFarland AP, Reynolds DA
    Blood, 2011;117(2):575-84.
    Species: Mouse
    Sample Types: Serum
  12. Phase 2 study of cladribine followed by rituximab in patients with hairy cell leukemia.
    Authors: Ravandi F, O'Brien S, Jorgensen J, Pierce S, Faderl S, Ferrajoli A, Koller C, Challagundla P, York S, Brandt M, Luthra R, Burger J, Thomas D, Keating M, Kantarjian H
    Blood, 2011;118(14):3818-23.
    Species: Human
    Sample Types: Serum
  13. Defucosylated humanized anti-CCR4 monoclonal antibody KW-0761 as a novel immunotherapeutic agent for adult T-cell leukemia/lymphoma.
    Authors: Ishii T, Ishida T, Utsunomiya A
    Clin. Cancer Res., 2010;16(5):1520-31.
    Species: Mouse
    Sample Types: Plasma
  14. Presence of IL-5 protein and IgE antibodies to staphylococcal enterotoxins in nasal polyps is associated with comorbid asthma.
    Authors: Bachert C, Zhang N, Holtappels G, De Lobel L, Van Cauwenberge P, Liu S, Lin P, Bousquet J, Van Steen K
    J. Allergy Clin. Immunol., 2010;126(5):962.
    Species: Human
    Sample Types: Tissue Homogenates
  15. Safety and clinical effect of subcutaneous human interleukin-21 in patients with metastatic melanoma or renal cell carcinoma: a phase I trial.
    Authors: Schmidt H, Brown J, Mouritzen U, Selby P, Fode K, Svane IM, Cook GP, Mollerup DH, Geertsen PF
    Clin. Cancer Res., 2010;16(21):5312-9.
    Species: Human
    Sample Types: Serum
  16. Effective treatment of a murine model of adult T-cell leukemia using depsipeptide and its combination with unmodified daclizumab directed toward CD25.
    Authors: Chen J, Zhang M, Ju W, Waldmann TA
    Blood, 2009;113(6):1287-93.
    Species: Mouse
    Sample Types: Serum
  17. Urinary proinflammatory cytokine response in renal transplant recipients with polyomavirus BK viruria.
    Authors: Sadeghi M, Daniel V, Schnitzler P, Lahdou I, Naujokat C, Zeier M, Opelz G
    Transplantation, 2009;88(9):1109-16.
    Species: Human
    Sample Types: Urine
  18. Smoking, inflammatory patterns and postprandial hypertriglyceridemia.
    Authors: Kabagambe EK, Ordovas JM, Tsai MY, Borecki IB, Hopkins PN, Glasser SP, Arnett DK
    Atherosclerosis, 2009;203(2):633-9.
    Species: Human
    Sample Types: Serum
  19. An open-label, two-arm, phase I trial of recombinant human interleukin-21 in patients with metastatic melanoma.
    Authors: Davis ID, Skrumsager BK, Cebon J, Nicholaou T, Barlow JW, Moller NP, Skak K, Lundsgaard D, Frederiksen KS, Thygesen P, McArthur GA
    Clin. Cancer Res., 2007;13(12):3630-6.
    Species: Human
    Sample Types: Serum
  20. 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 Types: Serum
  21. Expression and release of soluble HLA-E is an immunoregulatory feature of endothelial cell activation.
    Authors: Coupel S, Moreau A, Hamidou M, Horejsi V, Soulillou JP, Charreau B
    Blood, 2007;109(7):2806-14.
    Species: Human
    Sample Types: Serum
  22. Short communication: decreasing soluble CD30 and increasing IFN-gamma plasma levels are indicators of effective highly active antiretroviral therapy.
    Authors: Sadeghi M, Susal C, Daniel V, Naujokat C, Zimmermann R, Huth-Kuhne A, Opelz G
    AIDS Res. Hum. Retroviruses, 2007;23(7):886-90.
    Species: Human
    Sample Types: Plasma
  23. Effective treatment of a murine model of adult T-cell leukemia using 211At-7G7/B6 and its combination with unmodified anti-Tac (daclizumab) directed toward CD25.
    Authors: Zhang Z, Zhang M, Garmestani K, Talanov VS, Plascjak PS, Beck B, Goldman C, Brechbiel MW, Waldmann TA
    Blood, 2006;108(3):1007-12.
    Species: Human
    Sample Types: Serum
  24. Effective therapy for a murine model of human anaplastic large-cell lymphoma with the anti-CD30 monoclonal antibody, HeFi-1, does not require activating Fc receptors.
    Authors: Zhang M, Yao Z, Zhang Z, Garmestani K, Goldman CK, Ravetch JV, Janik J, Brechbiel MW, Waldmann TA
    Blood, 2006;108(2):705-10.
    Species: Human
    Sample Types: Serum
  25. Activating Fc receptors are required for antitumor efficacy of the antibodies directed toward CD25 in a murine model of adult t-cell leukemia.
    Authors: Zhang M, Zhang Z, Garmestani K, Goldman CK, Ravetch JV, Brechbiel MW, Carrasquillo JA, Waldmann TA
    Cancer Res., 2004;64(16):5825-9.
    Species: Human
    Sample Types: Serum
  26. Thymus and activation-regulated chemokine (TARC/CCL17) produced by mouse epidermal Langerhans cells is upregulated by TNF-alpha and IL-4 and downregulated by IFN-gamma.
    Authors: Xiao T, Fujita H, Saeki H, Mitsui H, Sugaya M, Tada Y, Kakinuma T, Torii H, Nakamura K, Asahina A, Tamaki K
    Cytokine, 2003;23(4):126-32.
    Species: Human
    Sample Types: Serum
  27. Comparative analysis of CA125, tissue polypeptide specific antigen, and soluble interleukin-2 receptor alpha levels in sera, cyst, and ascitic fluids from patients with ovarian carcinoma.
    Authors: Sedlaczek P, Frydecka I, Gabrys M, Van Dalen A, Einarsson R, Harlozinska A
    Cancer, 2002;95(9):1886-93.
    Species: Human
    Sample Types: Ascites Fluid
  28. 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 Types: Serum
  29. Medium-dose UVA1 cold-light phototherapy in the treatment of severe atopic dermatitis.
    Authors: von Kobyletzki G, Pieck C, Hoffmann K, Freitag M, Altmeyer P
    J. Am. Acad. Dermatol., 1999;41(6):931-7.
    Species: Human
    Sample Types: Serum

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