| Solution Information | help | |
| Enzyme: | Interleukin-8 | |
| inhibitor: | BDBM62334 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Protocol: 1. MCF7 cells stably overexpressing NOD2 2. MDP-LD (Invivogen) 3. IL-8 OptEIA ELISA set (BD Biosciences, San Diego, CA) 4. SpectraMax 190 spectrophotometer (Molecular Devices) Protocol: 1. Compounds were diluted in DMEM 10% FBS plus Pen/Strep (culture medium), and 10 uL were added into respective wells (96-well culture plate) to reach desired final concentrations. 2. MDP was added, in a final concentration of 5.4 ug/ml, to a culture medium suspension of MCF-7 cells (3,6x10+5 cells per mililiter). 3. Pre-induced MCF-7 cells were added into the respective wells (140 ul per well) and kept at 37oC (5% CO2 incubator) for 24 hours. 4. Thirty microliters of culture medium was used to measure IL-8 production using an IL-8 ELISA kit from BD according to the manufacturer's protocol. Briefly, 96-well ELISA plates (BD Biosciences) were coated with 100 uL per well of Capture Antibody diluted in Coating Buffer (100 mM Sodium carbonate, pH9.5) for overnight at 4oC. The wells were w | |
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