Rat IL-2 Quantikine ELISA Kit

(16 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Specificity
    Natural and recombinant rat IL-2
  • 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.
Product Summary
The Quantikine Rat IL-2 Immunoassay is a 4.5 hour solid phase ELISA designed to measure rat IL-2 levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant rat IL-2 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural rat IL-2 showed dose response 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 rat IL-2.

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
Sample123123
n202020202020
Mean13047814911394951550
Standard Deviation4.210.531.913.830.885.4
CV%3.22.22.19.96.25.5

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=8) 110 93-120
EDTA Plasma (n=4) 102 90-117
Heparin Plasma (n=4) 100 92-116
Serum (n=14) 96 83-114
Linearity
To assess the linearity of the assay, four or more samples containing and/or spiked with various concentrations of rat IL-2 were diluted with Calibrator Diluent and then assayed.
Rat IL-2 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-2
IL-2 (Interleukin 2) is a critical cytokine in T cell biology. It promotes T cell activation and expansion, the development, maintenance and function of regulatory T cells (Treg), and the differentiation of CD8+ T cells into terminal effector cells and memory cells. IL-2 signals through a receptor complex composed of CD25/IL-2 R alpha, IL-2 R beta, and the Common gamma Chain (gamma c). IL-2 R beta is also a component of the IL-15 receptor complex. Gamma c is also a signaling subunit in the receptors for IL-4, -7, -9, -15, and -21.
    • Long Name
      Interleukin 2
    • Entrez Gene IDs
      3558 (Human); 16183 (Mouse); 116562 (Rat); 396868 (Porcine); 280822 (Bovine); 403989 (Canine); 100034204 (Equine); 751114 (Feline); 100302458 (Rabbit);
    • Alternate Names
      aldesleukin; IL2; IL-2; IL-2lymphokine; interleukin 2; interleukin-2; involved in regulation of T-cell clonal expansion; T cell growth factor; T-cell growth factor; TCGF;
    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. 50 µL Standard, Control, or Sample
    6.   Add 50 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 4 times for a total of 5 washes.

    8. 100 µL Conjugate
    9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    10.   Aspirate and wash 5 times.

    11. 100 µL Substrate Solution
    12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    13. 100 µL Stop Solution
    14.   Add 100 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.

    16 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Melatonin synergized with cyclosporine A improves cardiac allograft survival by suppressing inflammation and apoptosis.
      Authors: Liu C, Hong T, Shao M, Chen Z, Wang C
      Mol Med Rep, 2014;10(3):1323-8.
      Species: Rat
      Sample Type: Serum
    2. Oridonin promotes CD4+/CD25+ Treg differentiation, modulates Th1/Th2 balance and induces HO-1 in rat splenic lymphocytes.
      Authors: Hu AP, Du JM, Li JY, Liu JW
      Inflamm. Res., 2008;57(4):163-70.
      Species: Rat
      Sample Type: Cell Culture Supernates
    3. Molecular identification of lyso-glycerophosphocholines as endogenous immunosuppressives in bovine and rat gonadal fluids.
      Authors: Foulds LM, Boysen RI, Crane M, Yang Y, Muir JA, Smith AI, de Kretser DM, Hearn MT, Hedger MP
      Biol. Reprod., 2008;79(3):525-36.
      Species: Rat
      Sample Type: Cell Culture Supernates
    4. Regulation of liver cytochrome P450 by activation of brain dopaminergic system: physiological and pharmacological implications.
      Authors: Wojcikowski J, Golembiowska K, Daniel WA
      Biochem. Pharmacol., 2008;76(2):258-67.
      Species: Rat
      Sample Type: Plasma
    5. Suppression of autoimmune retinal disease by lovastatin does not require Th2 cytokine induction.
      Authors: Gegg ME, Harry R, Hankey D, Zambarakji H, Pryce G, Baker D, Adamson P, Calder V, Greenwood J
      J. Immunol., 2005;174(4):2327-35.
      Species: Rat
      Sample Type: Cell Culture Supernates
    6. Chlorpromazine and loxapine reduce interleukin-1beta and interleukin-2 release by rat mixed glial and microglial cell cultures.
      Authors: Labuzek K, Kowalski J, Gabryel B, Herman ZS
      Eur Neuropsychopharmacol, 2005;15(1):23-30.
      Species: Rat
      Sample Type: Cell Culture Supernates
    7. Oral (gavage), in utero and post-natal exposure of Sprague-Dawley rats to low doses of tributyltin chloride. Part II: effects on the immune system.
      Authors: Tryphonas H, Cooke G, Caldwell D, Bondy G, Parenteau M, Hayward S, Pulido O
      Food Chem. Toxicol., 2004;42(2):221-35.
      Species: Rat
      Sample Type: Serum
    8. Gene transfer of tumor necrosis factor inhibitor improves the function of lung allografts.
      Authors: Tagawa T, Kozower BD, Kanaan SA, Daddi N, Muraoka M, Oka T, Ritter JH, Patterson GA
      J. Thorac. Cardiovasc. Surg., 2004;127(6):1558-63.
      Species: Rat
      Sample Type: Tissue Homogenates
    9. Insulin treatment improves hepatic morphology and function through modulation of hepatic signals after severe trauma.
      Authors: Schubert T, Horch RE
      Ann. Surg., 2004;240(2):340-9.
      Species: Rat
      Sample Type: Tissue Homogenates
    10. Insulin attenuates the systemic inflammatory response in endotoxemic rats.
      Authors: Jeschke MG, Klein D, Bolder U, Einspanier R
      Endocrinology, 2004;145(9):4084-93.
      Species: Rat
      Sample Type: Tissue Homogenates
    11. Recipient intramuscular cotransfection of naked plasmid transforming growth factor beta1 and interleukin 10 ameliorates lung graft ischemia-reperfusion injury.
      Authors: D&amp;amp;apos;Ovidio F
      J. Thorac. Cardiovasc. Surg., 2002;124(2):259-69.
      Species: Rat
      Sample Type: BALF
    12. Suppression of in vivo tumorigenicity of rat hepatoma cell line KDH-8 cells by soluble TGF-beta receptor type II.
      Authors: Zhao W, Kobayashi M, Ding W, Yuan L, Seth P, Cornain S, Wang J, Okada F, Hosokawa M
      Cancer Immunol. Immunother., 2002;51(7):381-8.
      Species: Rat
      Sample Type: Cell Culture Supernates
    13. Rat peripheral CD4+CD8+ T lymphocytes are partially immunocompetent thymus-derived cells that undergo post-thymic maturation to become functionally mature CD4+ T lymphocytes.
      Authors: Jimenez E, Sacedon R, Vicente A, Hernandez-Lopez C, Zapata AG, Varas A
      J. Immunol., 2002;168(10):5005-13.
      Species: Rat
      Sample Type: Cell Culture Supernates
    14. Melatonin fails to modulate immune parameters influenced by calorie restriction in aging Fischer 344 rats.
      Authors: Pahlavani MA, Vargas DA, Evans TR, Shu JH, Nelson JF
      Exp. Biol. Med. (Maywood), 2002;227(3):201-7.
      Species: Rat
      Sample Type: Cell Culture Supernates
    15. Local anticandidal immune responses in a rat model of vaginal infection by and protection against Candida albicans.
      Authors: de Bernardis F, Santoni G, Boccanera M, Spreghini E, Adriani D, Morelli L, Cassone A
      Infect. Immun., 2000;68(6):3297-304.
      Species: Rat
      Sample Type: Vaginal Fluid
    16. Cloning and characterization of a retroviral plasmid, pCC1, for combination suicide gene therapy.
      Authors: Kuiper M, Sanches R, Gaken JA, Bignon YJ
      BioTechniques, 2000;28(3):572-4, 576.
      Species: Rat
      Sample Type: Cell Culture Supernates
    Expand to show all 16 Citations
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