hh Rat IFN-gamma Quantikine ELISA Kit RIF00: R&D Systems

Rat IFN-gamma Quantikine ELISA Kit

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

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
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean86.9215115982.82051135
Standard Deviation3.54.423814.548
CV%4229.77.14.2

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 108 89-119
Serum (n=6) 100 89-111
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of rat IFN-gamma in each matrix were diluted with Calibrator Diluent and then assayed.
Rat IFN-gamma Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IFN-gamma
IFN-gamma (Interferon-gamma) is the prototype proinflammatory cytokine and is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells. It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, upregulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects. In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation. IFN-gamma dimers signal through a receptor complex of two IFN-gamma R1 and two IFN-gamma R2 subunits.
    • Long Name
      Interferon gamma
    • Entrez Gene IDs
      3458 (Human); 15978 (Mouse); 25712 (Rat); 396991 (Porcine); 281237 (Bovine); 403801 (Canine); 493965 (Feline);
    • Alternate Names
      IFG; IFI; IFNG; IFNgamma; IFN-gamma; Immune interferon; interferon gamma; interferon, gamma;
    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. 120 µL Conjugate
    9.   Add 120 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.

    21 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Biological effects of bone marrow mesenchymal stem cells on hepatitis B virus in�vitro
      Authors: WP Zheng, BY Zhang, ZY Shen, ML Yin, Y Cao, HL Song
      Mol Med Rep, 2017;15(5):2551-2559.
      Species: Rat
      Sample Type: Cell Culture Supernates
    2. Potassium channels Kv1.3 and KCa3.1 cooperatively and compensatorily regulate antigen-specific memory T cell functions
      Authors: EY Chiang, T Li, S Jeet, I Peng, J Zhang, WP Lee, J DeVoss, P Caplazi, J Chen, S Warming, DH Hackos, S Mukund, CM Koth, JL Grogan
      Nat Commun, 2017;8(0):14644.
      Species: Rat
      Sample Type: Cell Culture Supernates
    3. Melatonin attenuates the TLR4-mediated inflammatory response through MyD88- and TRIF-dependent signaling pathways in an in vivo model of ovarian cancer.
      Authors: Chuffa L, Fioruci-Fontanelli B, Mendes L, Ferreira Seiva F, Martinez M, Favaro W, Domeniconi R, Pinheiro P, Delazari Dos Santos L, Martinez F
      BMC Cancer, 2015;15(0):34.
      Species: Rat
      Sample Type: Tissue Culture Supernates
    4. Iron-hepcidin dysmetabolism, anemia and renal hypoxia, inflammation and fibrosis in the remnant kidney rat model.
      Authors: Garrido P, Ribeiro S, Fernandes J, Vala H, Bronze-da-Rocha E, Rocha-Pereira P, Belo L, Costa E, Santos-Silva A, Reis F
      PLoS ONE, 2015;10(4):e0124048.
      Species: Rat
      Sample Type: Serum
    5. 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
    6. Comprehensive gene expression profiling of rat lung reveals distinct acute and chronic responses to cigarette smoke inhalation.
      Authors: Stevenson CS, Docx C, Webster R, Battram C, Hynx D, Giddings J, Cooper PR, Chakravarty P, Rahman I, Marwick JA, Kirkham PA, Charman C, Richardson DL, Nirmala NR, Whittaker P, Butler K
      Am. J. Physiol. Lung Cell Mol. Physiol., 2007;293(5):L1183-93.
      Species: Rat
      Sample Type: BALF
    7. Changes of gene expression of iron regulatory proteins during turpentine oil-induced acute-phase response in the rat.
      Authors: Sheikh N, Dudas J, Ramadori G
      Lab. Invest., 2007;87(7):713-25.
      Species: Rat
      Sample Type: Serum
    8. The age-related attenuation in long-term potentiation is associated with microglial activation.
      Authors: Griffin R, Nally R, Nolan Y, McCartney Y, Linden J, Lynch MA
      J. Neurochem., 2006;99(4):1263-72.
      Species: Rat
      Sample Type: Tissue Homogenates
    9. 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
    10. Immunotherapy of murine prostate cancer using whole tumour cells killed ex vivo by cytosine deaminase/5-fluorocytosine suicide-gene therapy.
      Authors: Pandha HS, Cook D, Greenhalgh R, Dalgleish A
      BJU Int., 2005;95(9):1336-43.
      Species: Rat
      Sample Type: Cell Culture Supernates
    11. Effects of pyrrolidine dithiocarbamate, an NF-kappaB inhibitor, on cytokine expression and ocular inflammation in experimental autoimmune anterior uveitis.
      Authors: Fang IM, Yang CH, Lin CP, Yang CM, Chen MS
      J Ocul Pharmacol Ther, 2005;21(2):95-106.
      Species: Rat
      Sample Type: Aqueous humor
    12. Interleukin-10 aerosol reduces proinflammatory mediators in bronchoalveolar fluid of endotoxemic rat.
      Authors: Hofstetter C, Flondor M, Hoegl S, Muhl H, Zwissler B
      Crit. Care Med., 2005;33(10):2317-22.
      Species: Rat
      Sample Type: BALF
    13. Peroxisome proliferator-activated receptor-gamma ligands ameliorate experimental autoimmune myocarditis associated with inhibition of self-sensitive T cells.
      Authors: Yuan Z, Liu Y, Liu Y, Zhang J, Kishimoto C, Ma A, Liu Z
      J. Cardiovasc. Pharmacol., 2004;43(6):868-75.
      Species: Rat
      Sample Type:
    14. Ameliorating effects of the immunomodulator 3-(2-ethylphenyl)-5-(3-methoxyphenyl)-1H-1,2,4-triazole in an experimental model of colitis in the rat.
      Authors: Stasi MA, Ruggiero V, Ursillo A, Taurelli R, Agliano M, Weber E, Lorenzoni P, Sorrentino V, Pacifici L, Carminati P
      Eur. J. Pharmacol., 2004;494(2):263-72.
      Species: Rat
      Sample Type: Tissue Homogenates
    15. Alveolar macrophage cytotoxicity for normal lung fibroblasts is mediated by nitric oxide release.
      Authors: Morgan DL, Shines CJ
      Toxicol In Vitro, 2004;18(1):139-46.
      Species: Rat
      Sample Type: Cell Culture Supernates
    16. Salivary gland genetic vaccination: a scalable technology for promoting distal mucosal immunity and heightened systemic immune responses.
      Authors: Tucker SN, Lin K, Stevens S, Scollay R, Bennett MJ, Olson DC
      Vaccine, 2004;22(19):2500-4.
      Species: Rat
      Sample Type: Cell Culture Supernates
    17. 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
    18. 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
    19. 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
    20. Chimeric smooth muscle-specific enhancer/promoters: valuable tools for adenovirus-mediated cardiovascular gene therapy.
      Authors: Ribault S, Neuville P, Mechine-Neuville A, Auge F, Parlakian A, Gabbiani G, Paulin D, Calenda V
      Circ. Res., 2001;88(5):468-75.
      Species: Rat
      Sample Type: Cell Culture Supernates
    21. Role of IL-18 in acute lung inflammation.
      Authors: Jordan JA, Guo RF, Yun EC, Sarma V, Warner RL, Crouch LD, Senaldi G, Ulich TR, Ward PA
      J. Immunol., 2001;167(12):7060-8.
      Species: Rat
      Sample Type: BALF
    Expand to show all 21 Citations
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