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Assay Type
Quantitative Sandwich ELISA Development Module
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Assay Format
96-well microplate*
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Assay Length
3 hours 35 mins to 4 hours 50 mins**
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Sample Type
Whole Cells
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Sufficient Materials
For five 96-well microplates***
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Specificity
Please see the
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
* 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.
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.
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
MODULE CONTENTS
- Capture Antibody
- Biotinylated-conjugated Detection Antibody
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
Trademarks and registered trademarks are the property of their respective companies.
Preparation and Storage
Background: IL-4
IL-4 (Interleukin-4) is a cytokine that is primarily expressed by Th2-biased CD4+ T cells, mast cells, basophils, and eosinophils. It promotes cell proliferation, survival, and immunoglobulin class switch to IgG4 and IgE in human B cells, acquisition of the Th2 phenotype by naïve CD4+ T cells, priming and chemotaxis of mast cells, eosinophils, and basophils, and the proliferation and activation of epithelial cells. IL-4 plays a dominant role in the development of allergic inflammation and asthma. The type I receptor for IL-4, which is expressed on hematopoietic cells, is a heterodimer of the ligand binding IL-4 R? and the common ? chain (a shared subunit of the receptors for IL-2, -7, -9, -15, and -21). The type II receptor on nonhematopoietic cells consists of IL-4 R? and IL-13 R?1. The type II receptor also transduces IL-13 mediated signals.
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Entrez Gene IDs:
3565 (Human); 16189 (Mouse); 287287 (Rat); 397225 (Porcine); 280824 (Bovine); 403785 (Canine); 574281 (Primate); 100302454 (Rabbit);
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Long Name:
Interleukin 4
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Aliases:
B cell growth factor 1; BCDF; B-cell stimulatory factor 1; BCGF1; BCGF-1; binetrakin; BSF1; BSF-1; IL4; IL-4; IL-4B_cell stimulatory factor 1; interleukin 4; interleukin-4; Lymphocyte stimulatory factor 1; MGC79402; pitrakinra