Akademik Veri Yönetim Sistemi
EUROPEAN CYTOKINE NETWORK, cilt.25, sa.2, ss.30-34, 2014 (SCI-Expanded)
Measurements of cytokines in cell culture supernatants are widely used to evaluate the immune response. Cytokine levels in secretomes are usually quantified using enzyme-linked immunosorbent assays (ELISA), which have easy, sensitive, specific, rapid, cost-effective, and reproducible protocols. To our knowledge, the stability of cytokines in secretomes has not been hitherto investigated. We present data that involve; time-dependent changes during storage at +4 degrees C, and the effects of freeze-thaw cycles in samples frozen at -80 degrees C, instant freezing of samples with liquid nitrogen, and addition of protease inhibitors on the stability of certain cytokines (TNF-alpha, MIP-2, IFN-gamma, IL-6, IL-10, IL-17A), in secrotomes of spleen and lymph nodes from tumor-bearing animals. Our results show that IL-6 remains stable, MIP-2, IFN-gamma and IL-10 are somewhat stable, while TNF-alpha and IL-17A are degradable cytokines: instant freezing by liquid nitrogen or adding protease inhibitor does not preserve the stability of these cytokines. From these results it can be concluded that, if possible, TNF-alpha measurements should be perform in fresh samples, and IL-17A and IL-10 samples can be stored at -80 degrees C, but should be used at the first thaw.