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| ID | Components | Size |
| RSP001-C01 | Pre-coated Anti-IFN-γ Antibody Microplate | 1 plate |
| RSP001-C02 | Positive Stimulus | 10 μg |
| RSP001-C03 | Biotin-Anti-IFN-γ Antibody | 50 μL |
| RSP001-C04 | Streptavidin-HRP | 50 μL |
| RSP001-C05 | Washing Buffer (10x) | 50 mL |
| RSP001-C06 | Dilution Buffer (1x) | 50 mL |
| RSP001-C07 | AEC Dilution | 25 mL |
| RSP001-C08 | AEC Solution A (20x) | 0.8 mL |
| RSP001-C09 | AEC Solution B (20x) | 0.8 mL |
| RSP001-C10 | AEC Solution C (20x) | 0.8 mL |
This kit is used to detect IFN-γ at the cellular level.
It is for research use only.
Your experiment will include 6 simple steps:
a) A monoclonal antibody specific for human IFN-γ has been pre-coated onto a polyvinylidene difluoride (PVDF)-backed microplate.
b) 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.
c) After washing away any cells and unbound substances, a biotinylated antibody specific for human IFN-γ is added to the wells. Following a wash to remove any unbound biotinylated antibody, HRP conjugated to streptavidin is added.
d) Wash the plate and add the Streptavidin-HRP diluted by Dilution Buffer to the plate.
e) Wash the plate and add AEC. A reddish-brown colored precipitate forms and appears as spots at the sites of cytokine localization, with each individual spot representing an individual IFN-γ secreting cell.
f) SThe spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.
5x10⁴ cells/well PBMC were stimulated with 0.2 μg/mL Positive Stimulus and cultured at 37℃ and 5% CO2 for 20 h, 328 spots were produced. The following example data is for reference only.
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