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According to globally recognized standards of humane endpoints, animals were euthanized if they lost more than 20% of their body weight, had ulceration or necrosis at the tumor site, were unable to get food or water, were extremely lethargic, or displayed symptoms of pain or distress like hunching, a rough coat, or difficulty breathing

Merck Life Science N.V, Sigma, Netherlands, #NS02L-1ML) in PBS for 30 min

This is a critical part of Phase II detoxification

[DOI] [PubMed] [Google Scholar] 309.Ivanov B.N., Borisova-Mubarakshina M.M., Kozuleva M.A

This includes reduced inflammation, increased fibroblast activity, and restoration of ECM proteins

Researchers assume refrigeration alone is sufficient, but light exposure and repeated freeze-thaw cycles compound degradation risk