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 for Human IL-13 ELISpot Kit
- 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 for Human IL-13 ELISpot Kit
- 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
