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Feline IFN-gamma DuoSet ELISA

Catalog #: DY764
9 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY764
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Feline IFN-gamma ELISA Standard Curve
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Product Details
Procedure
Citations (9)
FAQs
Supplemental Products
Reviews
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Feline IFN-gamma DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
62.5 - 4,000 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 feline 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

Block Buffer: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Reagent Diluent: 0.1% BSA, 0.05% Tween 20 in Tris-buffered Saline (20 mM Trizma base, 150 mM NaCI) 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

N/A Feline IFN-gamma ELISA Standard Curve View Larger

Feline IFN-gamma ELISA Standard Curve

Product Datasheets

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Product Datasheet
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SDS

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 of Block Buffer 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 Feline 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.

9 Citations: Showing 1 - 9
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  1. Mucosal Immune Response to Feline Enteric Coronavirus Infection
    Authors: M Pearson, A LaVoy, S Evans, A Vilander, C Webb, B Graham, E Musselman, J LeCureux, S VandeWoude, GA Dean
    Viruses, 2019-09-27;11(10):.
    Species: Feline
    Sample Types: Cell Culture Supernates
  2. Mechanisms utilized by feline adipose-derived mesenchymal stem cells to inhibit T lymphocyte proliferation
    Authors: N Taechangam, SS Iyer, NJ Walker, B Arzi, DL Borjesson
    Stem Cell Res Ther, 2019-06-25;10(1):188.
    Species: Feline
    Sample Types: Cell Culture Supernates
  3. Activation of upper respiratory tract mucosal innate immune responses in cats by liposomal toll-like receptor ligand complexes delivered topically
    Authors: W Wheat, L Chow, J Coy, E Contreras, M Lappin, S Dow
    J. Vet. Intern. Med., 2019-02-15;0(0):.
    Species: Feline
    Sample Types: Cell Culture Supernates
  4. Human and feline adipose-derived mesenchymal stem cells have comparable phenotype, immunomodulatory functions, and transcriptome
    Authors: KC Clark, FA Fierro, EM Ko, NJ Walker, B Arzi, CG Tepper, H Dahlenburg, A Cicchetto, A Kol, L Marsh, WJ Murphy, N Fazel, DL Borjesson
    Stem Cell Res Ther, 2017-03-20;8(1):69.
    Species: Feline
    Sample Types: Cell Culture Supernates
  5. Identification and genotyping of feline infectious peritonitis-associated single nucleotide polymorphisms in the feline interferon-gamma gene.
    Authors: Hsieh L, Chueh L
    Vet Res, 2014-05-21;45(0):57.
    Species: Feline
    Sample Types: Plasma
  6. Production of IFN-gamma in feline whole blood after incubation with potential T-cell epitopes of the nucleocapsid protein of feline coronavirus.
    Authors: Rossi G, Cornaro C, Battilani M, Pocacqua V, Paltrinieri S
    Vet. Microbiol., 2011-02-13;150(3):248-56.
    Species: Feline
    Sample Types: Plasma
  7. Development and validation of a multiplex microsphere-based assay for detection of domestic cat (Felis catus) cytokines.
    Authors: Wood BA, O'Halloran KP, Vandewoude S
    Clin. Vaccine Immunol., 2011-01-05;18(3):387-92.
    Species: Feline
    Sample Types: Cell Culture Supernates
  8. feG-COOH blunts eosinophilic airway inflammation in a feline model of allergic asthma.
    Authors: Declue AE, Schooley E, Nafe LA, Reinero CR
    Inflamm. Res., 2009-03-03;58(8):457-62.
    Species: Feline
    Sample Types: BALF
  9. A recombinant subunit vaccine formulation protects against lethal Nipah virus challenge in cats.
    Authors: McEachern JA, Bingham J, Crameri G, Green DJ, Hancock TJ, Middleton D, Feng YR, Broder CC, Wang LF, Bossart KN
    Vaccine, 2008-06-02;26(31):3842-52.
    Species: Feline
    Sample Types: Plasma

FAQs

  1. Is PBS a suitable alternative with which to formulate the Reagent Diluent?

    • Our lab has determined that Tris-buffered saline generates optimal assay performance in comparison to PBS for this assay. This assay is validated with Reagent Diluent formulated using Tris-buffered saline.

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