Canine IFN-gamma ELISpot Kit

Catalog # Availability Size / Price Qty
PEL781
XEL781
EL781
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Citations (10)
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Canine IFN-gamma ELISpot Kit Summary

Assay Type
Quantitative Sandwich ELISA
Format
96-well PVDF-backed microplate
Assay Length
3 hours 15 mins to 4 hours 30 mins*
Sample Type
Whole Cells
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.


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 canine 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 canine 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 canine 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 canine 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 canine 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

  • Canine IFN-gamma Microplate
  • Biotinylated Detection Antibody
  • Streptavidin conjugated to Alkaline Phosphatase
  • Dilution Buffers
  • Wash Buffer Concentrate
  • BCIP/NBT Chromogen
  • Canine 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

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
⚠ WARNING: This product can expose you to chemicals including methanol, which is known to the State of California to cause reproductive toxicity with developmental effects. For more information, go to www.P65Warnings.ca.gov.

Citations for Canine 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.

10 Citations: Showing 1 - 10
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  1. Cas9-specific immune responses compromise local and systemic AAV CRISPR therapy in multiple dystrophic canine models
    Authors: CH Hakim, SRP Kumar, DO Pérez-Lópe, NB Wasala, D Zhang, Y Yue, J Teixeira, X Pan, K Zhang, ED Million, CE Nelson, S Metzger, J Han, JA Louderman, F Schmidt, F Feng, D Grimm, BF Smith, G Yao, NN Yang, CA Gersbach, SJ Chen, RW Herzog, D Duan
    Nature Communications, 2021;12(1):6769.
    Species: Canine
    Sample Types: Whole Cells
  2. Short-term treatment of golden retriever muscular dystrophy (GRMD) dogs with rAAVrh74.MHCK7.GALGT2 induces muscle glycosylation and utrophin expression but has no significant effect on muscle strength
    Authors: PT Martin, DA Zygmunt, A Ashbrook, S Hamilton, D Packer, SM Birch, AK Bettis, CJ Balog-Alva, LJ Guo, PP Nghiem, JN Kornegay
    PLoS ONE, 2021;16(3):e0248721.
    Species: Canine
    Sample Types: Whole Cells
  3. An Entry-Triggering Protein of Ehrlichia Is a New Vaccine Candidate against Tick-Borne Human Monocytic Ehrlichiosis
    Authors: K Budachetri, O Teymournej, M Lin, Q Yan, M Mestres-Vi, GN Brock, Y Rikihisa
    MBio, 2020;11(4):.
    Species: Canine
    Sample Types: Whole Blood
  4. A DNA telomerase vaccine for canine cancer immunotherapy
    Authors: J Thalmensi, E Pliquet, C Liard, G Chamel, C Kreuz, T Bestetti, M Escande, A Kostrzak, AS Pailhes-Ji, E Bourges, M Julithe, L Bourre, O Keravel, P Clayette, T Huet, S Wain-Hobso, P Langlade-D
    Oncotarget, 2019;10(36):3361-3372.
    Species: Canine
    Sample Types: Cell Culture Supernates
  5. An Observational Study from Long-Term AAV Re-administration in Two Hemophilia Dogs
    Authors: J Sun, W Shao, X Chen, EP Merricks, L Wimsey, YL Abajas, GP Niemeyer, CD Lothrop, PE Monahan, RJ Samulski, TC Nichols, C Li
    Mol Ther Methods Clin Dev, 2018;10(0):257-267.
    Species: Canine
    Sample Types: Whole Cells
  6. Frequency of IFNgamma-producing T cells correlates with seroreactivity and activated T cells during canine Trypanosoma cruzi infection.
    Authors: Hartley A, Cooley G, Gwyn S, Orozco M, Tarleton R
    Vet Res, 2014;45(0):6.
    Species: Canine
    Sample Types: Whole Cells
  7. Microdystrophin ameliorates muscular dystrophy in the canine model of duchenne muscular dystrophy.
    Authors: Shin J, Pan X, Hakim C, Yang H, Yue Y, Zhang K, Terjung R, Duan D
    Mol Ther, 2013;21(4):750-7.
    Species: Canine
    Sample Types: Whole Cells
  8. Programmed death 1-mediated T cell exhaustion during visceral leishmaniasis impairs phagocyte function.
    Authors: Esch K, Juelsgaard R, Martinez P, Jones D, Petersen C
    J Immunol, 2013;191(11):5542-50.
    Species: Canine
    Sample Types: Whole Cells
  9. Long-term correction of inhibitor-prone hemophilia B dogs treated with liver-directed AAV2-mediated factor IX gene therapy.
    Authors: Niemeyer GP, Herzog RW, Mount J, Arruda VR, Tillson DM, Hathcock J, van Ginkel FW, High KA, Lothrop CD
    Blood, 2009;113(4):797-806.
    Species: Canine
    Sample Types: Whole Cells
  10. RNA-loaded CD40-activated B cells stimulate antigen-specific T-cell responses in dogs with spontaneous lymphoma.
    Authors: Mason NJ, Coughlin CM, Overley B, Cohen JN, Mitchell EL, Colligon TA, Clifford CA, Zurbriggen A, Sorenmo KU, Vonderheide RH
    Gene Ther., 2008;15(13):955-65.
    Species: Canine
    Sample Types: Whole Cells

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