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2.6. Quantitative analysis of cytokines with enzyme-linked immunosorbent assay (ELISA)

CC Canan Cakir-Aktas
EB Ebru Bodur
MY Muge Yemisci
KL Klaus van Leyen
HK Hulya Karatas
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Quantitative expression of inflammatory cytokines (IL-6, TNF-alpha, IL-1beta, IL-10, and TGF-beta) in the ischemic brain regions was detected by the sandwich ELISA method. IL-6 (Biolegend, United States), TNF-alpha (Biolegend, United States), IL-1beta (Thermo Fisher, Germany), IL-10 (Biolegend, United States), and TGF-beta (Biolegend, United States) were quantified with ELISA kits according to the manufacturer’s protocols. The levels of cytokines in naive, I/R-DMSO and I/R-ML351-treated brain samples were compared. Three mice in each group were analyzed for ELISA, and each sample was tested in duplicate. The absorbances of the samples were read at a wavelength of 532 nm on a microplate reader (SpectraMax M2, Molecular Devices, United States). The curve of the standard graph was employed to calculate the results. The results were normalized by the amount of total protein, which was determined by BCA assay in the samples (ng/mg protein).

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