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The bioactivity of IL12 was determined by measuring the amount of IFNγ produced by human T cells and mouse splenocytes. Anti-CD3–stimulated cells were diluted to 5 × 105 cells per mL, and 200 μL per well was added to a 96-well plate. The produced proteins were added to achieve a final concentration of 250 pmol/L. Cells cultured in media alone served as the control group. Following 72 hours of incubation, the supernatants were collected for cytokine quantification using sandwich ELISA, following the instructions provided by the manufacturer (DY285, R&D systems).
Copyright and License information: ©2024 The Authors; Published by the American Association for Cancer Research
This open access article is distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) license.
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