| Porcine IFN-gamma ELISpot Kit | |
| PART | DESCRIPTION |
| Microplate | 96-well PVDF-backed microplate coated with a monoclonala ntibody |
| Detection Antibody Concentrate | 120 uL of a 100X concentrated solution of biotinylated monoclonal antibody |
| Streptavidin-AP Concentrate A | 120 μL of a 100X concentrated solution of Streptavidin con jugated to Alkaline Phosphatase with preservatives |
| Dilution Buffer 1 | 12 mL of a buffer for diluting Detection Antibody Concentra te with preservatives. |
| Dilution Buffer 2 | 12 mL of a buffer for diluting Streptavidin-AP Concentrate A with preservatives. |
| Wash Buffer Concentrate | 20 mL of a 25X concentrated solution of a buffered surfact ant with preservative |
| Principle | The enzyme-linked immunospot (ELISpot) assay was originally developed for the detection of individual B cells secreting antigen-specific antibodies. This method has since been adapted for the detection of individual cells secreting specific cytokines or other antigens. ELISpot assays employ the quantitative sandwich enzyme-linked immunosorbent assay (ELISA) technique. A monoclonal antibody specific for porcine IFN-γ has been pre-coated onto a PVDF (polyvinylidene difluoride)-backed microplate.Appropriately stimulated cells are pipetted into the wells and the microplate is placed into a humidified 37 °C CO2 incubator for a specified period of time. During this incubation period, the immobilized antibody in the immediate vicinity of the secreting cells binds secreted IFN-γ. After washing away any cells and unbound substances, a biotinylated monoclonal antibody specific for porcine IFN-γ is added to the wells. Following a wash to remove any unbound biotinylated 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 appear as spots, with each individual spot representing an individual IFN-γ secreting cell. The spots can be counted with an ELISpot reader system or using a stereomicroscope. |
Porcine IFN-gamma ELISpot Kit
| Catalogue No | BSEK-330PO |
| Species | Porcine |
| Size | 96 T | 48 T |
| Price | POR |
For the quantitative determination of the frequency of cells releasing porcine Interferon gamma (IFN-γ).