Rat IFN-gamma ELISpot Kit

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XEL585
EL585
R&D Systems ELISpot and FluoroSpot Kits
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Citations (7)
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Rat 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 rat 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 rat 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 rat 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 rat 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 rat 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

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

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 Rat 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.

7 Citations: Showing 1 - 7
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  1. Effective immunotherapy of rat glioblastoma with prolonged intratumoral delivery of exogenous heat shock protein Hsp70.
    Authors: Shevtsov M, Pozdnyakov A, Mikhrina A, Yakovleva L, Nikolaev B, Dobrodumov A, Komarova E, Meshalkina D, Ischenko A, Pitkin E, Guzhova I, Margulis B
    Int J Cancer, 2014;135(9):2118-28.
    Species: Rat
    Sample Types: Whole Cells
  2. Imatinib ameliorates neuroinflammation in a rat model of multiple sclerosis by enhancing blood-brain barrier integrity and by modulating the peripheral immune response.
    Authors: Adzemovic M, Zeitelhofer M, Eriksson U, Olsson T, Nilsson I
    PLoS ONE, 2013;8(2):e56586.
    Species: Rat
    Sample Types: Whole Cells
  3. Combined Flt3L/TK gene therapy induces immunological surveillance which mediates an immune response against a surrogate brain tumor neoantigen.
    Authors: King GD, Muhammad AK, Larocque D
    Mol. Ther., 2011;19(10):1793-801.
    Species: Rat
    Sample Types: Whole Cells
  4. Steady-state migrating intestinal dendritic cells induce potent inflammatory responses in naive CD4+ T cells.
    Authors: Milling SW, Jenkins CD, Yrlid U, Cerovic V, Edmond H, McDonald V, Nassar M, Macpherson G
    Mucosal Immunol, 2009;2(2):156-65.
    Species: Rat
    Sample Types: Whole Cells
  5. Transition from enhanced T cell infiltration to inflammation in the myelin-degenerative central nervous system.
    Authors: Grundtner R, Dornmair K, Dahm R, Flugel A, Kawakami N, Zeitelhofer M, Schoderboeck L, Nosov M, Selzer E, Willheim M, Kiebler M, Wekerle H, Lassmann H, Bradl M
    Neurobiol. Dis., 2007;28(3):261-75.
    Species: Rat
    Sample Types: Whole Cells
  6. Comparative analysis of the fate of donor dendritic cells and B cells and their influence on alloreactive T cell responses under tacrolimus immunosuppression.
    Authors: Azhipa O, Kimizuka K, Nakao A, Toyokawa H, Okuda T, Neto JS, Alber SM, Kaizu T, Thomson AW, Demetris AJ, Murase N
    Clin. Immunol., 2005;114(2):199-209.
    Species: Rat
    Sample Types: Whole Cells
  7. The activation status of neuroantigen-specific T cells in the target organ determines the clinical outcome of autoimmune encephalomyelitis.
    Authors: Kawakami N, Lassmann S, Li Z, Odoardi F, Ritter T, Ziemssen T, Klinkert WE, Ellwart JW, Bradl M, Krivacic K, Lassmann H, Ransohoff RM, Volk HD, Wekerle H, Linington C, Flugel A
    J. Exp. Med., 2004;199(2):185-97.
    Species: Rat
    Sample Types: Whole Cells

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