Mouse IL-2 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 mouse IL-2. Complete ELISpot kits are ready-to-run and require no assay development or refinement.
This ELISpot assay employs a capture antibody specific for mouse IL-2, 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 mouse IL-2. 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 mouse IL-2 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 mouse IL-2
- 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
- Mouse IL-2 Microplate
- Biotinylated Detection Antibody
- Streptavidin conjugated to Alkaline Phosphatase
- Dilution Buffers
- Wash Buffer Concentrate
- BCIP/NBT Chromogen
- Mouse IL-2 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: IL-2
IL-2 (Interleukin 2) is a critical cytokine in T cell biology. It promotes T cell activation and expansion, the development, maintenance and function of regulatory T cells (Treg), and the differentiation of CD8+ T cells into terminal effector cells and memory cells. IL-2 signals through a receptor complex composed of CD25/IL-2 R alpha, IL-2 R beta, and the Common gamma Chain (gamma c). IL-2 R beta is also a component of the IL-15 receptor complex. Gamma c is also a signaling subunit in the receptors for IL-4, -7, -9, -15, and -21.
Citations for Mouse IL-2 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 - 6
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Extracellular overproduction of E7 oncoprotein of Iranian human papillomavirus type 16 by genetically engineered Lactococcus lactis
Authors: AH Mohseni, S Taghinezha, H Keyvani, V Razavilar
BMC Biotechnol., 2019-01-24;19(1):8.
Species: Mouse
Sample Types: Whole Cells
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Protection against human papillomavirus type 16-induced tumors in C57BL/6 mice by mucosal vaccination with Lactococcus lactis NZ9000 expressing E6 oncoprotein
Authors: S Taghinezha, AH Mohseni, H Keyvani, V Razavilar
Microb. Pathog., 2018-11-02;0(0):.
Species: Mouse
Sample Types: Whole Cells
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A whole recombinant yeast-based therapeutic vaccine elicits HBV X, S and Core specific T cells in mice and activates human T cells recognizing epitopes linked to viral clearance.
Authors: King T, Kemmler C, Guo Z, Mann D, Lu Y, Coeshott C, Gehring A, Bertoletti A, Ho Z, Delaney W, Gaggar A, Subramanian G, McHutchison J, Shrivastava S, Lee Y, Kottilil S, Bellgrau D, Rodell T, Apelian D
PLoS ONE, 2014-07-22;9(7):e101904.
Species: Mouse
Sample Types: Whole Cells
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HIV-1 adenoviral vector vaccines expressing multi-trimeric BAFF and 4-1BBL enhance T cell mediated anti-viral immunity.
Authors: Kanagavelu, Saravana, Termini, James M, Gupta, Sachin, Raffa, Francesc, Fuller, Katherin, Rivas, Yaelis, Philip, Sakhi, Kornbluth, Richard, Stone, Geoffrey
PLoS ONE, 2014-02-28;9(2):e90100.
Species: Mouse
Sample Types: Whole Cells
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Vaccination with a fusion protein that introduces HIV-1 gag antigen into a multitrimer CD40L construct results in enhanced CD8+ T cell responses and protection from viral challenge by vaccinia-gag.
Authors: Gupta, Sachin, Termini, James M, Raffa, Francesc, Williams, Cindi-An, Kornbluth, Richard, Stone, Geoffrey
J Virol, 2013-11-13;88(3):1492-501.
Species: Mouse
Sample Types: Whole Cells
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The critical role of kinase activity of interleukin-1 receptor-associated kinase 4 in animal models of joint inflammation.
Authors: Koziczak-Holbro M, Littlewood-Evans A, Pollinger B, Kovarik J, Dawson J, Zenke G, Burkhart C, Muller M, Gram H
Arthritis Rheum., 2009-06-01;60(6):1661-71.
Species: Mouse
Sample Types: Whole Cells
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