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Enzyme-linked immunosorbent assay (ELISA) assay

PZ Pengzhan Zhao
YW Yutian Wei
GS Guangchi Sun
LX Lei Xu
TW Tian Wang
YT Yufei Tian
HC Honglu Chao
YT Yiming Tu
JJ Jing Ji
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The brain tissues were collected from the mice after CCI 6 h. Briefly, added 5 mg of brain tissue to a tube along with 300 µL of extraction buffer and mix for 2 h at 4 °C with an electric homogenizer. Then, the brain tissues were centrifuged at 13,000 rpm for 20 min at 4 °C. The supernatant was extracted and diluted with sample buffer at a ratio of 1:1 and loaded onto the wells in duplicates. TNF-α, IL-6, IL-1β, and IL-10 expressions in the brain tissues were measured using a specific enzyme-linked immunosorbent assay (ELISA; R&D Systems Inc) according to the manufacturer’s instruction.

The conditioned medium was moved to a centrifuge tube and centrifuged at 1500 rpm for 10 min at 4 °C and samples were loaded onto the wells in duplicates. TNF-α, IL-6, IL-1β, and IL-10 expressions in culture supernatants were measured by a specific ELISA according to the manufacturer’s instruction.

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