Canine IFN-gamma ELISpot Kit

Catalog #: EL781
11 Citations Datasheet

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PEL781
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EL781

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Citations (11)

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Summary Product Datasheets Preparation and Storage Background

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

Product Datasheet

Preparation and Storage

Shipping

The product is shipped with polar packs. 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

⚠ 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.

11 Citations: Showing 1 - 10
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Efficacy and Immune Correlates of OMP-1B and VirB2-4 Vaccines for Protection of Dogs from Tick Transmission of Ehrlichia chaffeensis
Authors: K Budachetri, M Lin, RC Chien, W Zhang, GN Brock, Y Rikihisa
MBio, 2022-11-07;0(0):e0214022.
Species: Canine
Sample Types: Whole Cells
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, 2021-11-24;12(1):6769.
Species: Canine
Sample Types: Whole Cells
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, 2021-03-26;16(3):e0248721.
Species: Canine
Sample Types: Whole Cells
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, 2020-07-28;11(4):.
Species: Canine
Sample Types: Whole Blood
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, 2019-05-21;10(36):3361-3372.
Species: Canine
Sample Types: Cell Culture Supernates
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, 2018-08-04;10(0):257-267.
Species: Canine
Sample Types: Whole Cells
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, 2014-01-23;45(0):6.
Species: Canine
Sample Types: Whole Cells
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, 2013-10-23;191(11):5542-50.
Species: Canine
Sample Types: Whole Cells
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, 2013-01-15;21(4):750-7.
Species: Canine
Sample Types: Whole Cells
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, 2008-10-28;113(4):797-806.
Species: Canine
Sample Types: Whole Cells

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., 2008-03-13;15(13):955-65.
Species: Canine
Sample Types: Whole Cells