* Sold separately (see Other Reagents Required, below)
** Following an overnight incubation with capture antibody, 2 hour blocking step, and cell stimulation.
*** When using the protocol provided.

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PRODUCT SUMMARY
Complete are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. Kits are available for detection and enumeration of a single analyte or two analytes simultaneously. Complete ELISpot kits are ready-to-run and require no assay development or refinement. contain the basic components required to develop an ELISpot assay. They offer an economical alternative to buying separate antibodies.

ELISpot development modules are an alternative to ELISpot kits. A basic understanding of ELISpot assay development is required for the successful use of these reagents. Each investigator should optimize the coating conditions, the assay sensitivity, the type of enzyme and substrate, as well as the concentrations of the capture and detection antibodies to achieve desired results. The analyte-specific ELISpot Development Module and the contain the necessary components for analyte detection and visualization, respectively. These modules can be used together but are sold separately. Each module contains enough reagents for at least five 96-well microplates.

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PRODUCT FEATURES

• An economical alternative to ELISpot Kits
• Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
• Generic development protocols provide direction to start an optimization protocol
• Customize the assay to your specific needs

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MODULE CONTENTS

• Capture Antibody
• Biotinylated-conjugated Detection Antibody

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OTHER REAGENTS REQUIRED

• ELISpot Blue Color Module or equivalent (R&D Systems, Catalog # )
• PBS - 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2 - 7.4, 0.2 µm filtered
• Wash Buffer - 0.05% Tween^® 20 in PBS
• Blocking Buffer - 1% BSA, 5% Sucrose in PBS
• Reagent Diluent - 1% BSA in PBS, pH 7.2 - 7.4, 0.2 µm filtered
• 2 °C – 8 °C refrigerator
• 37 °C CO₂ incubator
• Positive Control - Use Recombinant or cells known to secrete
• 96-well plates - Nitrocellulose-bottom plates, PVDF-bottom Immunospot^® plates, or flat-bottom polystyrene Immulon^® ELISA plates
• Multi-channel pipette, squirt bottle, manifold dispenser, or automated microplate washer
• Dissection microscope or an automated ELISpot Reader
• Deionized H₂O

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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.