Murine ifn γ elisa development kit

Bloods were collected under terminal anaesthesia by cardiac puncture. Sera were prepared from infected and control mice (L. monocytogenes model) and stored at −80 °C. Murine IFN-γ ELISA Development Kit was used according to the manufacturer’s instructions (Peprotech). Nitric oxide production was measured using Griess Reagent kit (Promega, Wisconsin, USA).

The supernatants from mice cultured spleen cells used in the CFSE proliferation assay were evaluated for IFN-γ production using a Murine IFN-γ ELISA development kit (PEPROTECH, Mexico City, Mexico) following the instructions of manufacturer. Briefly, ELISA plates containing 1 μg/ml capture antibody per well were incubated at 4°C overnight. After addition of standards and samples, plates were incubated at room temperature for 2 h. The detection was performed with 0.5 μg/ml detection antibody. The plates were incubated at room temperature for 2 h. Avidin peroxidase diluted 1:2000 was added and incubated at room temperature for 30 min. A color reaction was developed using ABTS liquid substrate and absorbance values were measured at 405 nm using an ELISA plate reader.