Feline IFN-gamma ELISpot Kit
Feline IFN-gamma ELISpot Kit 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.
PRODUCT SUMMARY
ELISpot kits are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. This kit is designed for the detection and enumeration of feline IFN-gamma. Complete ELISpot kits are ready-to-run and require no assay development or refinement.
This ELISpot assay employs a capture antibody specific for feline IFN-gamma, pre-coated onto a PVDF-backed microplate. Appropriately stimulated cells are pipetted directly into the wells and the immobilized antibody in the immediate vicinity of the secreting cells binds secreted feline IFN-gamma. Following wash steps and incubation with a biotinylated detection antibody, alkaline-phosphatase conjugated to streptavidin is added. Unbound enzyme is subsequently removed by washing and a substrate solution (BCIP/NBT) is added. A blue-black colored precipitate forms at the sites of cytokine localization and appears as spots, with each individual spot representing an individual feline IFN-gamma secreting cell. The spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.
PRODUCT FEATURES
- Detect and quantitate individual cells secreting feline IFN-gamma
- High sensitivity - ELISpot assays can measure responses with frequencies well below 1 in 100,000 cells
- No in vitro expansion of cells required
- High-throughput - ELISpot assays use only a small number of primary cells
KIT CONTENTS
- Feline IFN-gamma Microplate
- Biotinylated Detection Antibody
- Streptavidin conjugated to Alkaline Phosphatase
- Dilution Buffers
- Wash Buffer Concentrate
- BCIP/NBT Chromogen
- Feline IFN-gamma Positive Control
OTHER REAGENTS REQUIRED
- Pipettes and pipette tips
- Deionized or distilled water
- Squirt bottle, manifold dispenser, or automated microplate washer
- 500 mL graduated cylinder
- 37 °C CO2 incubator
- Sterile culture media
- Dissection microscope or an automated ELISpot reader
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.
Citations for Feline IFN-gamma ELISpot Kit
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.
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Citations: Showing 1 - 4
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Modified-Live Feline Calicivirus Vaccination Elicits Cellular Immunity against a Current Feline Calicivirus Field Strain in an Experimental Feline Challenge Study
Authors: AM Spiri, M Novacco, ML Meli, M Stirn, B Riond, JE Fogle, FS Boretti, I Herbert, MJ Hosie, R Hofmann-Le
Viruses, 2021-08-31;13(9):.
Species: Feline
Sample Types: Whole Cells
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CD4(+)CD25(+) T regulatory cells inhibit CD8(+) IFN-gamma production during acute and chronic FIV infection utilizing a membrane TGF-beta-dependent mechanism.
Authors: Fogle JE, Mexas AM, Tompkins WA, Tompkins MB
AIDS Res. Hum. Retroviruses, 2010-02-01;26(2):201-16.
Species: Feline
Sample Types: Whole Cells
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Evaluation of ISCOM-adjuvanted subunit vaccines containing recombinant feline immunodeficiency virus Rev, OrfA and envelope protein in cats.
Authors: Huisman W, Schrauwen EJ, Pas SD, van Amerongen G, Rimmelzwaan GF, Osterhaus AD
Vaccine, 2008-04-01;26(21):2553-61.
Species: Feline
Sample Types: Whole Cells
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Cellular immune responses to feline immunodeficiency virus (FIV) induced by dual-subtype FIV vaccine.
Authors: Omori M, Pu R, Tanabe T, Hou W, Coleman JK, Arai M, Yamamoto JK
Vaccine, 2004-12-02;23(3):386-98.
Species: Feline
Sample Types: Whole Cells
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