Rat IFN-gamma DuoSet ELISA Summary
* 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 rat IFN-gamma. 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)
Scientific Data
Product Datasheets
Preparation and Storage
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.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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 Rat IFN-gamma 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.
18
Citations: Showing 1 - 10
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Allanblackia floribunda Seed Extract Attenuates the Ethanol-Induced Gastric Ulcer in Rats via the Inhibition of TNF-&alpha and INF-&gamma Levels and Modulation in the Expression of Ki67 Protein
Authors: FA Armah, IT Henneh, J Alake, W Ahlidja, B Amoani, EG Ofori, B Asante-Kye, GI Temitayo, CK Adokoh
BioMed Research International, 2021-01-11;2021(0):6694572.
Species: Rat
Sample Types: Serum
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The Poststroke Peripheral Immune Response Is Differentially Regulated by Leukemia Inhibitory Factor in Aged Male and Female Rodents
Authors: SM Davis, LA Collier, SJ Messmer, KR Pennypacke
Oxidative Medicine and Cellular Longevity, 2020-12-10;2020(0):8880244.
Species: Rat
Sample Types: Tissue Homogenates
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GABA potentiate the immunoregulatory effects of Lactobacillus brevis BGZLS10-17 via ATG5-dependent autophagy in vitro
Authors: SS Baji?, J ?oki?, M Dini?, S Tomi?, N Popovi?, E Brdari?, N Goli?, M Tolina?ki
Sci Rep, 2020-01-28;10(1):1347.
Species: Rat
Sample Types: Cell Culture Supernates
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An immunocompetent rat model of infection with Cryptosporidium species: Cryptosporidium hominis and Cryptosporidium parvum
Authors: DA Dayao, A Sheoran, A Carvalho, H Xu, G Beamer, G Widmer, S Tzipori
Int. J. Parasitol., 2019-11-22;0(0):.
Species: Rat
Sample Types: Cell Culture Supernates
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Cholinergic Stimulation by Pyridostigmine Bromide Before Myocardial Infarction Prevent Cardiac and Autonomic Dysfunction
Authors: CA Barboza, AR Fukushima, N Carrozzi, JF Machi, PMM Dourado, CT Mostarda, MC Irigoyen, L Nathanson, M Morris, EC Caperuto, B Rodrigues
Sci Rep, 2019-02-21;9(1):2481.
Species: Rat
Sample Types: Plasma
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Leukemia inhibitory factor modulates the peripheral immune response in a rat model of emergent large vessel occlusion
Authors: SM Davis, LA Collier, ED Winford, CC Leonardo, CT Ajmo, EA Foran, TJ Kopper, JC Gensel, KR Pennypacke
J Neuroinflammation, 2018-10-15;15(1):288.
Species: Rat
Sample Types: Tissue Homogenates
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Lactobacillus fermentum HP3-Mediated Fermented Hericium erinaceus Juice as a Health Promoting Food Supplement to Manage Diabetes Mellitus
Authors: C Chaiyasut, S Woraharn, BS Sivamaruth, N Lailerd, P Kesika, S Peerajan
J Evid Based Integr Med, 2018-01-01;23(0):2515690X18765.
Species: Rat
Sample Types: Serum
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Intestinal anti-inflammatory activity of Ground Cherry (Physalis angulata L.) standardized CO2 phytopharmaceutical preparation
Authors: LD Almeida Ju, AEV Quaglio, CAR de Almeida, LC Di Stasi
World J. Gastroenterol., 2017-06-28;23(24):4369-4380.
Species: Rat
Sample Types: Tissue Homogenates
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Impact of gestational nicotine exposure on intrauterine and fetal infection in a rodent model
Authors: M von Chamie, L Reyes, LF Hayward, MB Brown
Biol. Reprod., 2017-05-01;96(5):1071-1084.
Species: Rat
Sample Types: Whole Tissue
Applications: ELISA Capture -
Food-grade TiO2 impairs intestinal and systemic immune homeostasis, initiates preneoplastic lesions and promotes aberrant crypt development in the rat colon
Authors: S Bettini, E Boutet-Rob, C Cartier, C Com‚ra, E Gaultier, J Dupuy, N Naud, S Tach‚, P Grysan, S Reguer, N Thieriet, M R‚fr‚giers, D ThiaudiŠre, JP Cravedi, M CarriŠre, JN Audinot, FH Pierre, L Guzylack-P, E Houdeau
Sci Rep, 2017-01-20;7(0):40373.
Species: Rat
Sample Types: Tissue Homogenates
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Neuroprotection by Caffeine in Hyperoxia-Induced Neonatal Brain Injury
Authors: S Endesfelde, U Weichelt, E Strauá, A Schl”r, M Sifringer, T Scheuer, C Bhrer, T Schmitz
Int J Mol Sci, 2017-01-18;18(1):.
Species: Rat
Sample Types: Tissue Homogenates
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Impact of exercise training associated to pyridostigmine treatment on autonomic function and inflammatory profile after myocardial infarction in rats
Authors: Bruno Rodrigues
Int. J. Cardiol., 2016-10-26;0(0):.
Species: Rat
Sample Types: Plasma
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Expression of heme oxygenase-1 in neural stem/progenitor cells as a potential mechanism to evade host immune response.
Authors: Bonnamain V, Mathieux E, Thinard R, Thebault P, Nerriere-Daguin V, Leveque X, Anegon I, Vanhove B, Neveu I, Naveilhan P
Stem Cells, 2012-10-01;30(10):2342-53.
Species: Rat
Sample Types: Cell Culture Supernates
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Kinetics of IFN-gamma and IL-17 expression and production in active experimental autoimmune encephalomyelitis in Dark Agouti rats.
Authors: Momcilovic M, Miljkovic Z, Popadic D, Miljkovic D, Mostarica-Stojkovic M
Neurosci. Lett., 2008-10-07;447(2):148-52.
Species: Rat
Sample Types: Cell Culture Supernates
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Interaction between interferon gamma and insulin-like growth factor-1 in hippocampus impacts on the ability of rats to sustain long-term potentiation.
Authors: Maher FO, Clarke RM, Kelly A, Nally RE, Lynch MA
J. Neurochem., 2006-02-08;96(6):1560-71.
Species: Rat
Sample Types: Tissue Homogenates
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RDP58, a novel immunomodulatory peptide, ameliorates clinical signs of disease in the Lewis rat model of acute experimental autoimmune encephalomyelitis.
Authors: DeVry CG, Valdez M, Gao L, Wang J, Kotsch K, Volk HD, Bechmann I, Buelow R, Iyer S
J. Neuroimmunol., 2004-07-01;152(1):33-43.
Species: Rat
Sample Types: Tissue Homogenates
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Selective blockade of voltage-gated potassium channels reduces inflammatory bone resorption in experimental periodontal disease.
Authors: Valverde P, Kawai T, Taubman MA
J. Bone Miner. Res., 2004-01-01;19(1):155-64.
Species: Rat
Sample Types: Cell Culture Supernates
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A toxicokinetic study of nickel-induced immunosuppression in rats.
Authors: O'Donnell JM
Immunopharmacol Immunotoxicol, 2003-11-01;25(4):655-70.
Species: Mouse
Sample Types: Cell Culture Supernates
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