| Solution Information | help | |
| Enzyme: | Matrix metalloproteinase-14 [1-36] | |
| inhibitor: | BDBM48256 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | 1) HT1080 cells that had been stably transfected with the 7.2 kb 5'-flanking region of the MT1-MMP gene linked to the luciferase gene in the pGL3-Basic vector (Promega), were seeded into Greiner black, clear bottom plates (cat #781091). 7500 cells were seeded/well in 45ul of DMEM w/o phenol red supplemented with 10% FBS, 1mM Sodium Pyruvate, 2mM glutamine and 20ug/ml gentamicin. 2) Cells are left to adhere overnight at 37 Celsius degrees in a humid environment in the presence of 5% CO2. 3) After the overnight incubation, 5ul of diluted compound (final compound concentration is 10uM, final DMSO concentration is 1%) are added to appropriate wells. 4) 5ul of 10% DMSO are added to negative control wells. 5) 5ul of saturated sodium sulfate are added to positive control wells. 6) The cells are incubated with compound overnight at 37 Celsius degrees in a humid environment in the presence of 5% CO2. 7) After incubation 10ul of 16 fold diluted Alamar Blue (Biosource International DAL1100 | |
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