Rat IFN-gamma DuoSet ELISA

Catalog # Availability Size / Price Qty
DY585
Ancillary Products Available
Rat IFN-gamma  ELISA Standard Curve
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Product Details
Procedure
Citations (18)
FAQs
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Rat IFN-gamma DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
39.1 - 2,500 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* 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

Rat IFN-gamma ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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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  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. Neuroprotection by Caffeine in Hyperoxia-Induced Neonatal Brain Injury
    Authors: S Endesfelde, U Weichelt, E Strauá, A Schl”r, M Sifringer, T Scheuer, C Bhrer, T Schmitz
    Int J Mol Sci, 2017-01-18;18(1):.
    Species: Rat
    Sample Types: Tissue Homogenates
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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
  17. 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
  18. 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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