Dual color ifn γ il 4 elispot kit

Dual-Color IFN-γ/IL-4 ELISpot Kit (R&D Systems# ELD5217) was adopted for monitoring cross-protective responses of PMSB-inoculated hosts to CSC subsets. Briefly, thymocytes were harvested from host thymus 2 weeks after the boost. 5 × 10⁴ recipient thymocytes as responder cells and 3 × 10³ PMSB cells, or tumor spheroid cells as stimulators were respectively performed according to the manufacturer's protocol with common PSCs as stimulator control. Spots were automatically scanned and enumerated using ELISpot plate reader (Cellular Technology Ltd., Cleveland, OH).

The Dual-Color IFN-γ/IL-4 ELISpot Kit (R&D, USA ) was used to monitor the
primary and cross responses of immunized rats to TSA-treated and irradiated
NuTu-19 cells and tumor cells, respectively. Briefly, the rats were immunized
with TSA-treated and irradiated NuTu-19 cells as described above and sacrificed
2 weeks after the last boost vaccination. The spleen was collected before
splenocyte isolation. Assays using 3×10³ X-ray-irradiated tumor cells
as stimulator cells and 10⁵ recipient splenocytes as responder cells
were carried out. An ELISpot plate reader (CTL Analyzers; Cellular Technology,
USA) was used to automatically enumerate the spots for further analysis.