The concentration prostaglandin E2 in cell culture supernatants was measured by the Prostaglandin E2 EIA Kit—Monoclonal (Cayman Chemicals, Ann Arbor, MI, USA) following the manufacturer's instructions. The release of interferon-gamma and interferon-alpha by T cells cocultured with MEF was quantified using the Mouse IFN-gamma Platinum ELISA (eBioscience, San Diego, CA, USA) and VeriKine Mouse Interferon Alpha ELISA Kit (PBL Interferon Source, Piscataway, NJ, USA).
Mouse ifn gamma platinum elisa
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Mouse IFN-gamma Platinum ELISA is a laboratory equipment product used for the quantitative in vitro determination of mouse interferon-gamma (IFN-gamma) concentrations in cell culture supernatants, serum, and plasma.
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3 protocols using mouse ifn gamma platinum elisa
1
Quantifying Immune Cell Secretions
2
Cytokine Production by DNA-Vaccinated Splenocytes
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To assess capability of splenic lymphocytes of DNA vaccinated mice to produce cytokines when stimulated by T. gondii antigens, splenocytes were plated in Costar 24-well plates (Thermo Fisher Scientific, USA) at the density of 3.5×106 cells/well and stimulated with 50 µg/mL of STAg or 5 µg/mL of Con A (Sigma, USA) as positive control or media alone as negative control. After 72 hrs of incubation at 37°C in presence of 5% CO2, supernatants were collected in separate microtubes and centrifuged for 10 mins at 3000× rpm and then concentrations of IFN-γ and IL-10 were measured using commercial kits according to the manufacturer’s instructions (Mouse IFN-gamma Platinum ELISA and Mouse IL-10 Platinum ELISA; ebioscience, USA). All tests were repeated thrice.
3
Serum Insulin and IFN-gamma Analysis
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To analyze serum insulin concentrations, animals were fasted overnight and injected with 1 g/kg of D-glucose. Blood was isolated from the v. saphena using heparin coated plastics. Insulin was measured in plasma using the mouse ultrasensitive insulin ELISA kit (Alpco). IFN-g concentrations were measured with the mouse IFN-gamma Platinum ELISA (eBioscience) according to manufacturer's protocol. Plates were analyzed using a Mithras LB940 ELISA plate reader (Berthold technologies).
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