| Assay Method Information | |
| | Evaluation of the Ability to Inhibit TNF-α Secretion by human Resistin |
| Description: | Using THP-1 (human monocyte) cell line and ELISA system, the abilities to inhibit TNF-α secretion by hResistin (IC50) were evaluated for the compounds prepared in the above examples. The compounds target hResistin, and the abilities of the compounds to inhibit TNF-α secretion by human recombinant Resistin in human monocyte (THP-1) were evaluated. The abilities to inhibit TNF-α secretion were assessed by ELISA quantifying the amount of antibody with the enzyme as a marker using an antigen-antibody reaction.Specifically, the cultured cells were spun down at 1,500 rpm for 2 minutes, the supernatant was removed and then the cells were re-suspended in 10 mL of complete RPMI-1640 medium. After counting the number of cells using Luna Automated Cell Counter, cells were plated into 96-well assay plates at 50 μL per well. The cells cultured for 24 hours were treated with the test substances in accordance with the concentration for 1 hour, and then the supernatant was collected and subjected to ELISA assay. In the ELISA assay, the absorbance at 450 nm (OD450 nm) was measured using Flexstation 3. |
| Affinity data for this assay | |
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