Human IL-13 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 human IL-13. Complete ELISpot kits are ready-to-run and require no assay development or refinement.
This ELISpot assay employs a capture antibody specific for human IL-13, 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 human IL-13. 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 human IL-13 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 human IL-13
- 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
- Human IL-13 Microplate
- Biotinylated Detection Antibody
- Streptavidin conjugated to Alkaline Phosphatase
- Dilution Buffers
- Wash Buffer Concentrate
- BCIP/NBT Chromogen
- Human IL-13 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-13
IL-13 (Interleukin-13) plays a key role in the pathogenesis of allergy, cancer, and tissue fibrosis. It is secreted by several helper T cell subsets, NK cells, mast cells, eosinophils, basophils, and visceral smooth muscle cells. IL-13 suppresses the production of proinflammatory cytokines and other cytotoxic substances by macrophages, fibroblasts, and endothelial cells. It promotes B cell activation, immunoglobulin class switching to IgE, and the upregulation of CD23/Fc epsilon RII. IL-13 signals through a receptor complex containing IL-13 R alpha 1 and IL-4 R alpha. This complex also functions as the type 2 IL-4 receptor. IL-13 R alpha 2 binds IL-13 and prevents IL-13 from signaling through the IL-13 R alpha 1/IL-4 R alpha complex. IL-13-bound IL-13 R alpha 2 can directly promote tumor cell invasiveness and the development of tissue fibrosis.
Citations for Human IL-13 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 - 3
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Helminth antigens modulate immune responses in cells from multiple sclerosis patients through TLR2-dependent mechanisms.
Authors: Correale J, Farez M
J. Immunol., 2009-10-07;183(9):5999-6012.
Species: Human
Sample Types: Whole Cells
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A multidonor ELISPOT study of IL-1 beta, IL-2, IL-4, IL-6, IL-13, IFN-gamma and TNF-alpha release by cryopreserved human peripheral blood mononuclear cells.
Authors: Bailey T, Stark S, Grant A, Hartnett C, Tsang M, Kalyuzhny A
J. Immunol. Methods, 2002-12-15;270(2):171-82.
Species: Human
Sample Types: Whole Cells
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Eosinophils express functional IL-13 in eosinophilic inflammatory diseases.
Authors: Schmid-Grendelmeier P, Altznauer F, Fischer B, Bizer C, Straumann A, Menz G, Blaser K, Wuthrich B, Simon HU
J. Immunol., 2002-07-15;169(2):1021-7.
Species: Human
Sample Types: Whole Cells
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