@Article{ecn.2014.0353,
AUTHOR = {Ozbey G, Gorczynski R, Erin N},
TITLE = {Stability of cytokines in supernatants of stimulated mouse immune cells},
JOURNAL = {European Cytokine Network},
VOLUME = {25},
YEAR = {2014},
NUMBER = {2},
PAGES = {30--34},
URL = {http://www.techscience.com/ECN/v25n2/65560},
ISSN = {1952-4005},
ABSTRACT = {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℃, and the effects of freeze-thaw cycles in samples frozen at -80℃, instant freezing of samples
with liquid nitrogen, and addition of protease inhibitors on the stability of certain cytokines (TNF-α, MIP-2, IFN-γ,
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-γ and IL-10 are somewhat stable, while TNF-α 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-α measurements should be perform in fresh
samples, and IL-17A and IL-10 samples can be stored at -80℃, but should be used at the first thaw.},
DOI = {10.1684/ecn.2014.0353}
}