Porcine 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 porcine 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 porcine 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 porcine 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 porcine 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 porcine 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
- Porcine IFN-gamma Microplate
- Biotinylated Detection Antibody
- Streptavidin conjugated to Alkaline Phosphatase
- Dilution Buffers
- Wash Buffer Concentrate
- BCIP/NBT Chromogen
- Porcine 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 Porcine 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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The role of zinc sulfate in enhancing cellular and humoral immune responses to foot-and-mouth disease vaccine
Authors: Ko, MK;Kim, HW;Park, SH;Park, JH;Kim, SM;Lee, MJ;
Virus research
Species: Porcine
Sample Types: Cell Culture Supernates
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Using a concurrent challenge with porcine circovirus 2 and porcine reproductive and respiratory syndrome virus to compare swine vaccination programs
Authors: A Madapong, K Saeng-Chut, A Tantituvan, D Nilubol
Scientific Reports, 2022-09-15;12(1):15524.
Species: Porcine
Sample Types: Cell Culture Supernates
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A chimeric porcine reproductive and respiratory syndrome virus (PRRSV)-2 vaccine is safe under international guidelines and effective both in experimental and field conditions
Authors: HY Choi, SH Lee, SH Ahn, JC Choi, JY Jeong, BJ Lee, YL Kang, SS Hwang, JK Lee, SW Lee, SY Park, CS Song, IS Choi, JB Lee
Research in Veterinary Science, 2021-01-23;135(0):143-152.
Species: Porcine
Sample Types: Cell Culture Supernates
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Evaluation of the recombinant proteins RlpB and VacJ as a vaccine for protection against Glaesserella parasuis in pigs
Authors: SJ Hau, SL Luan, CL Loving, TL Nicholson, J Wang, SE Peters, D Seilly, LA Weinert, PR Langford, AN Rycroft, BW Wren, DJ Maskell, AW Tucker, SL Brockmeier
BMC Vet. Res., 2020-05-27;16(1):167.
Species: Porcine
Sample Types: Whole Cells
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Cell-mediated immune response and protective efficacy of porcine reproductive and respiratory syndrome virus modified-live vaccines against co-challenge with PRRSV-1 and PRRSV-2
Authors: A Madapong, K Saeng-Chut, A Boonsoongn, A Tantituvan, D Nilubol
Sci Rep, 2020-02-03;10(1):1649.
Species: Porcine
Sample Types: Whole Cells
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Simultaneous deletion of the 9GL and UK genes from the African swine fever virus Georgia 2007 isolate offers increased safety and protection against homologous challenge.
Authors: O'Donnell V, Risatti G, Holinka L, Krug P, Carlson J, Velazquez-Salinas L, Azzinaro P, Gladue D, Borca M
J Virol, 2016-12-16;0(0):.
Species: Porcine
Sample Types: Whole Cells
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Association of the Host Immune Response with Protection Using a Live Attenuated African Swine Fever Virus Model.
Authors: Carlson J, O'Donnell V, Alfano M, Velazquez Salinas L, Holinka L, Krug P, Gladue D, Higgs S, Borca M
Viruses, 2016-10-22;8(10):.
Species: Porcine
Sample Types: Whole Cells
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In Vivo Validation of Predicted and Conserved T Cell Epitopes in a Swine Influenza Model
PLoS ONE, 2016-07-13;11(7):e0159237.
Species: Porcine
Sample Types: Whole Cells
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Cationic lipid-formulated DNA vaccine against hepatitis B virus: immunogenicity of MIDGE-Th1 vectors encoding small and large surface antigen in comparison to a licensed protein vaccine.
Authors: Endmann A, Klunder K, Kapp K, Riede O, Oswald D, Talman E, Schroff M, Kleuss C, Ruiters M, Juhls C
PLoS ONE, 2014-07-03;9(7):e101715.
Species: Porcine
Sample Types: Whole Cells
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Elicitation of broad CTL response against HIV-1 by the DNA vaccine encoding artificial multi-component fusion protein MultiHIV--study in domestic pigs.
Authors: Molder T, Adojaan M, Kaldma K, Ustav M, Sikut R
Vaccine, 2009-10-29;28(2):293-8.
Species: Porcine
Sample Types: Whole Cells
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