Human IL-8/CXCL8 ELISpot Development Module, 5 Plate
Human IL-8/CXCL8 ELISpot Development Module, 5 Plate 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.
Complete ELISpot kits 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. ELISpot Development Modules 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 ELISpot Blue Color Module 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.
- 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
- Human IL-8/CXCL8 Capture Antibody
- Human IL-8/CXCL8 Biotinylated-conjugated Detection Antibody
OTHER REAGENTS REQUIRED
- ELISpot Blue Color Module or equivalent (R&D Systems, Catalog # SEL002)
- 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 Human IL-8/CXCL8 or cells known to secrete Human IL-8/CXCL8
- 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
Interleukin-8 (IL-8), also known as CXCL8, is a chemokine that is upregulated at sites of inflammation where it promotes neutrophil infiltration and activation. IL-8 can form homodimers and heterodimers with CXCL4/PF4. Its bioactivity is regulated by proteolytic truncations, citrullination, and the decoy receptor DARC. IL-8 signals through CXCR1/IL-8 RA, which is also used by CXCL6, and through CXCR2/IL-8 RB, which is used by multiple CXC chemokines. IL-8 also binds to Serpin A1/alpha-1 Antitrypsin which prevents IL-8 interaction with CXCR1.
Citation for Human IL-8/CXCL8 ELISpot Development Module, 5 Plate
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.
1 Citation: Showing 1 - 1
ELISpot analysis of LPS-stimulated leukocytes: human granulocytes selectively secrete IL-8, MIP-1beta and TNF-alpha.
Authors: Smedman C, Gardlund B, Nihlmark K, Gille-Johnson P, Andersson J, Paulie S
J. Immunol. Methods, 2009;346(1):1-8.
Sample Types: Whole Cells
Are mouse or rat CXCL8/IL-8 products offered?
Mouse and rat do not have a gene which encodes CXCL8/IL-8. In mouse, the functional homologs to human CXCL8/IL-8 are CXCL1/KC and CXCL2/MIP-2. In rat, the functional homolog to CXCL8/human IL-8 is CXCL3/CINC-2.
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